Sulfonamide inhibitors of aspartyl protease

ABSTRACT

The present invention relates to a novel class of sulfonamides which are aspartyl protease inhibitors. In one embodiment, this invention relates to a novel class of HIV aspartyl protease inhibitors characterized by specific structural and physicochemical features. This invention also relates to pharmaceutical compositions comprising these compounds. The compounds and pharmaceutical compositions of this invention are particularly well suited for inhibiting HIV-1 and HIV-2 protease activity and consequently, may be advantageously used as anti-viral agents against the HIV-1 and HIV-2 viruses. This invention also relates to methods for inhibiting the activity of HIV aspartyl protease using the compounds of this invention and methods for screening compounds for anti-HIV activity.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.10/094,763, filed Mar. 8, 2002 now U.S. Pat. No. 6,720,335, the entiredisclosure of which is hereby incorporated by reference; which is adivisional of U.S. patent application Ser. No. 09/409,808, filed Sep.30, 1999 now U.S. Pat. No. 6,392,046, which is a divisional of U.S.patent application Ser. No. 09/115,394, filed Jul. 14, 1998, now U.S.Pat. No. 5,977,137; which is a divisional of U.S. patent applicationSer. No. 08/393,460 filed Feb. 23, 1995, now U.S. Pat. No. 5,783,701;which is a continuation-in-part of U.S. patent application Ser. No.08/142,327, filed Nov. 24, 1993, now U.S. Pat. No. 5,585,397; which is acontinuation-in-part of U.S. patent application Ser. No. 07/941,982,filed Sep. 8, 1992, now abandoned.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a novel class of sulfonamides which areaspartyl protease inhibitors. In one embodiment, this invention relatesto a novel class of HIV aspartyl protease inhibitors characterized byspecific structural and physicochemical features. This invention alsorelates to pharmaceutical compositions comprising these compounds. Thecompounds and pharmaceutical compositions of this invention areparticularly well suited for inhibiting HIV-1 and HIV-2 proteaseactivity and consequently, may be advantageously used as anti-viralagents against the HIV-1 and HIV-2 viruses. This invention also relatesto methods for inhibiting the activity of HIV aspartyl protease usingthe compounds of this invention and methods for screening compounds foranti-HIV activity.

BACKGROUND OF THE INVENTION

The human immunodeficiency virus (“HIV”) is the causative agent foracquired immunodeficiency syndrome (“AIDS”)—a disease characterized bythe destruction of the immune system, particularly of CD4⁺ T-cells, withattendant susceptibility to opportunistic infections—and its precursorAIDS-related complex (“ARC”)—a syndrome characterized by symptoms suchas persistent generalized lymphadenopathy, fever and weight loss.

As in the case of several other retroviruses, HIV encodes the productionof a protease which carries out post-translational cleavage of precursorpolypeptides in a process necessary for the formation of infectiousvirions (S. Crawford et al., “A Deletion Mutation in the 5′ Part of thepol Gene of Moloney Murine Leukemia Virus Blocks Proteolytic Processingof the gag and pol Polyproteins”, J. Virol., 53, p. 899 (1985)). Thesegene products include pol, which encodes the virion RNA-dependent DNApolymerase (reverse transcriptase), an endonuclease, HIV protease, andqag, which encodes the core-proteins of the virion (H. Toh et al.,“Close Structural Resemblance Between Putative Polymerase of aDrosophila Transposable. Genetic Element 17.6 and pol gene product ofMoloney Murine Leukemia Virus”, EMBO J., 4, p. 1267 (1985); L. H. Pearlet al., “A Structural Model for the Retroviral Proteases”, Nature, pp.329-351 (1987); M. D. Power et al., “Nucleotide Sequence of SRV-1, aType D Simian Acquired Immune Deficiency Syndrome Retrovirus”, Science,231, p. 1567 (1986)).

A number of synthetic anti-viral agents have been designed to targetvarious stages in the replication cycle of HIV. These agents includecompounds which block viral binding to CD4⁺ T-lymphocytes (for example,soluble CD4), and compounds which interfere with viral replication byinhibiting viral reverse transcriptase (for example, didanosine andzidovudine (AZT)) and inhibit integration of viral DNA into cellular DNA(M. S. Hirsh and R. T. D'Aqulia, “Therapy for Human ImmunodeficiencyVirus Infection”, N. Eng. J. Med., 328, p. 1686 (1993)). However, suchagents, which are directed primarily to early stages of viralreplication, do not prevent the production of infectious virions inchronically infected cells. Furthermore, administration of some of theseagents in effective amounts has led to cell-toxicity and unwanted sideeffects, such as anemia and bone marrow suppression.

More recently, the focus of anti-viral drug design has been to createcompounds which inhibit the formation of infectious virions byinterfering with the processing of viral polyprotein precursors.Processing of these precursor proteins requires the action ofvirus-encoded proteases which are essential for replication (Kohl, N. E.et al. “Active HIV Protease is Required for Viral Infectivity” Proc.Natl. Acad. Sci. USA, 85, p. 4686 (1988)). The anti-viral potential ofHIV protease inhibition has been demonstrated using peptidal inhibitors.Such peptidal compounds, however, are typically large and complexmolecules that tend to exhibit poor bioavailability and are notgenerally consistent with oral administration. Accordingly, the needstill exists for compounds that can effectively inhibit the action ofviral proteases, for use as agents for preventing and treating chronicand acute viral infections.

SUMMARY OF THE INVENTION

The present invention provides a novel class of compounds, andpharmaceutically acceptable derivatives thereof, that are useful asinhibitors of aspartyl proteases, in particular, HIV aspartyl protease.These compounds can be used alone or in combination with othertherapeutic or prophylactic agents, such as anti-virals, antibiotics,immunomodulators or vaccines, for the treatment or prophylaxis of viralinfection.

According to a preferred embodiment, the compounds of this invention arecapable of inhibiting HIV viral replication in human CD₄ ⁺ T-cells.These compounds are useful as therapeutic and prophylactic agents totreat or prevent infection by HIV-1 and related viruses which may resultin asymptomatic infection, AIDS-related complex (“ARC”), acquiredimmunodeficiency syndrome (“AIDS”), or similar disease of the immunesystem.

It is a principal object of this invention to provide a novel class ofsulfonamides which are aspartyl protease inhibitors, and particularly,HIV aspartyl protease inhibitors. This novel class of sulfonamides isrepresented by formula I:

wherein:

A is selected from the group consisting of H; Het; —R¹-Het; —R¹—C₁-C₆alkyl, which may be optionally substituted with one or more groupsselected from the group consisting of hydroxy, C₁-C₄ alkoxy, Het,—O-Het, —NR²—CO—N(R²)(R²) and —CO—N(R²) (R²); and —R¹—C₂-C₆ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het, —O-Het,—NR²—CO—N(R²)(R²) and —CO—N(R²)(R²);

each R¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)— and—NR²—C(O)—C(O)—;

each Het is independently selected from the group consisting of C₃-C₇cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturatedor unsaturated heterocycle, containing one or more heteroatoms selectedfrom N, N(R²), O, S and S(O)_(n), wherein said heterocycle mayoptionally be benzofused; and wherein any member of said Het may beoptionally substituted with one or more substituents selected from thegroup consisting of oxo, —OR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R²,—C(O)—N(R²) (R²), —S(O)₂—N(R²)(R²), —N(R²)—C(O)—R², —C(O)—R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—Ar, methylenedioxy, —N(R²)—S(O)₂(R²),halo, —CF₃, —NO₂, Ar and —O—Ar;

each R² is independently selected from the group consisting of H andC₁-C₃ alkyl optionally substituted with Ar;

B, when present, is —N(R²)—C(R³)(R³)—C(O)—;

x is 0 or 1;

each R³ is independently selected from the group consisting of H, Het,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl,wherein any member of said R³, except H, may be optionally substitutedwith one or more substituents selected from the group consisting of—OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Het, —CN, —SR², —CO₂R²,NR²—C(O)—R²;

each n is independently 1 or 2;

D and D′ are independently selected from the group consisting of Ar;C₁-C₄ alkyl, which may be optionally substituted with one or more groupsselected from C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₂-C₄ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar;C₃-C₆ cycloalkyl, which may be optionally substituted with or fused withAr; and C₅-C₆ cycloalkenyl, which may be optionally substituted with orfused with Ar;

each Ar is independently selected from the group consisting of phenyl;3-6 membered carbocyclic ring and 5-6 membered heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) andN(R²), wherein said carbocyclic or heterocyclic ring may be saturated orunsaturated and optionally substituted with one or more groups selectedfrom the group consisting of oxo, —OR², —R², —N(R²)(R²), —N(R²)—C(O)—R²,—R²—OH, —CN, —CO₂R², —C(O)—N(R²)(R²), halo and —CF₃;

E is selected from the group consisting of Het; O-Het; Het-Het; —O—R³;—NR²R³; C₁-C₆ alkyl, which may be optionally substituted with one ormore groups selected from the group consisting of R⁴ and Het; C₂-C₆alkenyl, which may be optionally substituted with one or more groupsselected from the group consisting of R⁴ and Het; C₃-C₆ saturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and C₅-C₆ unsaturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and

each R⁴ is independently selected from the group consisting of —OR²,—C(O)—NHR², —S(O)₂—NHR², halo, —NR²—C(O)—R² and —CN.

It is a also an object of this invention to provide pharmaceuticalcompositions comprising the sulfonamides of formula I and methods fortheir use as inhibitors of HIV aspartyl protease.

It is a further object of this invention to provide a novel class of HIVaspartyl protease inhibitor compounds characterized by the followingnovel combination of structural and physicochemical features:

(1) a first and a second hydrogen bond acceptor moiety, at least one ofwhich is more highly polarizable than a carbonyl, said moieties beingthe same or different, and being capable of hydrogen bonding with thehydrogen atoms of the flap water molecule of an HIV aspartyl proteasewhen the compound is bound thereto;

(2) substantially hydrophobic moieties which associate with the P₁ andP₁′ binding pockets of said HIV aspartyl protease when the compound isbound thereto;

(3) a third hydrogen bonding moiety, which may be either a hydrogen bonddonor or acceptor, capable of simultaneously hydrogen bonding to Asp25and Asp25′ of said HIV aspartyl protease when the compound is boundthereto;

(4) an additional occupied volume of space of at least 100 Å³ when thecompound is bound to the active site of said HIV aspartyl protease, saidspace overlapping with the volume of space that would be filled by anative substrate of said HIV aspartyl protease or a nonhyrolyzableisostere thereof;

(5) a deformation energy of binding of the compound to said HIV aspartylprotease of not greater than 10 kcal/mole; and

(6) a neutral or favorable enthalpic contribution from the sum of allelectrostatic interactions between the compound and the protease whenthe compound is bound to said HIV aspartyl protease.

It is also an object of this invention to provide pharmaceuticalcompositions comprising compounds having the above-mentioned featuresand methods for their use as inhibitors of HIV aspartyl protease.

It is a further object of this invention to provide a method foridentification, design, or prediction of HIV aspartyl proteaseinhibitors comprising the steps of:

-   -   (a) selecting a candidate compound of defined chemical structure        containing a first and a second hydrogen bond acceptor moiety,        at least one of which is more highly polarizable than a        carbonyl, said moieties being the same or different; a third        hydrogen bonding moiety, which may be either a hydrogen bond        donor or acceptor; and at least two substantially hydrophobic        moieties;    -   (b) determining a low-energy conformation for binding of said        compound to the active site of an HIV aspartyl protease;    -   (c) evaluating the capability of said first and second hydrogen        bond acceptor moieties to form hydrogen bonds to the flap water        molecule of said HIV aspartyl protease when said compound is        bound thereto in said conformation;    -   (d) evaluating the capability of said substantially hydrophobic        moieties to associate with the P₁ and P₁′ binding pockets of        said HIV aspartyl protease when said compound is bound thereto        in said conformation;    -   (e) evaluating the capability of said third hydrogen bonding        moiety to form hydrogen bonds to Asp25 and Asp25′ of said HIV        aspartyl protease when said compound is bound thereto in said        conformation;    -   (f) evaluating the overlap of the occupied volume of said        compound when said compound is bound to said HIV aspartyl        protease in said conformation and the occupied volume of a        native substrate of HIV aspartyl protease or a nonhydrolyzable        isostere thereof, when said polypeptide is bound to said HIV        aspartyl protease;    -   (g) evaluating the deformation energy of binding of said        compound to said HIV aspartyl protease;    -   (h) evaluating the enthalpic contribution of the sum of all        electrostatic interactions between said compound and said HIV        aspartyl protease when said compound is bound thereto in said        conformation; and    -   (i) accepting or rejecting said candidate compound as an HIV        protease inhbitor based upon the determinations and evaluations        carried out in steps (b) through (h).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a stereo drawing of a low-energy conformation of Compound140, as predicted by computer-modelling.

FIG. 2 depicts a stereo drawing of the actual crystal structure ofCompound 140, as observed by X-ray crystallography.

FIG. 3 depicts a stereo drawing of the correlation between the predicted(thin line) and observed (thick line) conformation of Compound 140.

DETAILED DESCRIPTION OF THE INVENTION

In order that the invention herein described may be more fullyunderstood, the following detailed description is set forth. In thedescription, the following abbreviations are used:

Designation Reagent or Fragment Ac acetyl Me methyl Et ethyl Bzl benzylTrityl triphenylmethyl Asn D- or L-asparagine Ile D- or L-isoleucine PheD- or L-phenylalanine Val D- or L-valine Boc tert-butoxycarbonyl Cbzbenzyloxycarbonyl (carbobenzyloxy) Fmoc 9-fluorenylmethoxycarbonyl DCCdicyclohexylcarbodiimide DIC diisopropylcarbodiimide EDC1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride HOBt1-hydroxybenzotriazole HOSu 1-hydroxysuccinimide TFA trifluoroaceticacid DIEA diisopropylethylamine DBU 1,8-diazabicyclo(5.4.0)undec-7-eneEtOAc ethyl acetate

The following terms are employed herein:

Unless expressly stated to the contrary, the terms “—SO₂—” and “—S(O)₂—”as used herein refer to a sulfone or sulfone derivative (i.e., bothappended groups linked to the S), and not a sulfinate ester.

For the compounds of formula I, and intermediates thereof, thestereochemistry of the explicitly shown hydroxyl is defined relative toD on the adjacent carbon atom, when the molecule is drawn in an extendedzig-zag representation (such as that drawn for compounds of formula XI,XV, XXII, XXIII and XXXI). If both OH and D reside on the same side ofthe plane defined by the extended backbone of the compound, thestereochemistry of the hydroxyl will be referred to as “syn”. If OH andD reside on opposite sides of that plane, the stereochemistry of thehydroxyl will be referred to as “anti”.

The term “heterocyclic” refers to a stable 5-7 membered monocycle or8-11 membered bicyclic heterocycle which is either saturated orunsaturated, and which may be optionally benzofused if monocyclic. Eachheterocycle consists of carbon atoms and from one to four heteroatomsselected from the group consisting of nitrogen, oxygen and sulfur. Asused herein, the terms “nitrogen and sulfur heteroatoms” include anyoxidized form of nitrogen and sulfur, and the quaternized form of anybasic nitrogen. The heterocyclic ring may be attached by any heteroatomof the cycle which results in the creation of a stable structure.Preferred heterocycles defined above include, for example,benzimidazolyl, imidazolyl, imidazolinoyl, imidazolidinyl, quinolyl,isoquinolyl, indolyl, pyridyl, pyrrolyl, pyrrolinyl, pyrazolyl,pyrazinyl, quinoxolyl, piperidinyl, morpholinyl, thiamorpholinyl, furyl,thienyl, triazolyl, thiazolyl, β-carbolinyl, tetrazolyl, thiazolidinyl,benzofuanoyl, thiamorpholinyl sulfone, benzoxazolyl, oxopiperidinyl,oxopyrroldinyl, oxoazepinyl, azepinyl, isoxazolyl, tetrahydropyranyl,tetrahydrofuranyl, thiadiazoyl, benzodioxolyl, thiophenyl,tetrahydrothiophenyl and sulfolanyl.

The terms “HIV protease” and “HIV aspartyl protease” are usedinterchangeably and refer to the aspartyl protease encoded by the humanimmunodeficiency virus type 1 or 2. In a preferred embodiment of thisinvention, these terms refer to the human immunodeficiency virus type 1aspartyl protease.

The term “hydrophobic” refers to a moiety which tends not to dissolvereadily in water and is often fat-soluble. Hydrophobic moieties include,but are not limited to, hydrocarbons, such as alkanes, alkenes, alkynes,cycloalkanes, cycloalkenes, cycloalkynes and aromatic hydrocarbons, suchas aryls, certain saturated and unsaturated heterocycles and moietiesthat are substantially similar to the side chains of hydrophobic naturaland unnatural α-amino acids, including valine, leucine, isoleucine,methionine, phenylalanine, α-amino isobutyric acid, alloisoleucine,tyrosine, and tryptophan.

The term “substantially hydrophobic” refers to a hydrophobic moietywhich may optionally contain polar atoms or groups in the region of themoiety which are solvent exposed when the compound is bound in theactive site of an aspartyl protease.

The term “linker moiety” refers to a group within a compound, said groupconsisting of a backbone of 1-6 atoms selected from the group consistingof C, N, O, S and P, said backbone being substituted with, fused to orotherwise associated with a substantially hydrophobic group capable ofassociating with the P₁ or P₁′ binding pocket of an HIV aspartylprotease when said compound is bound thereto. In alternative embodimentsof this invention, such linker moieties may optionally be substitutedwith a group or groups which occupy a volume of space overlapping withthe volume of space that would be filled by a native substrate of HIVaspartyl protease or a nonhydrolyzable isostere thereof.

The term “more highly polarizable than a carbonyl” refers to a moietyhaving a polarizability (α) greater than that of a carbonyl group of acorresponding aldehyde, ketone, ester or amide moiety.

The term “pharmaceutically effective amount” refers to an amounteffective in treating HIV infection in a patient. The term“prophylactically effective amount” refers to an amount effective inpreventing HIV infection in a patient. As used herein, the term“patient” refers to a mammal, including a human.

The term “pharmaceutically acceptable carrier or adjuvant” refers to anon-toxic carrier or adjuvant that may be administered to a patient,together with a compound of this invention, and which does not destroythe pharmacological activity thereof.

As used herein, the compounds of this invention, including the compoundsof formula I, are defined to include pharmaceutically acceptablederivatives thereof. A “pharmaceutically acceptable derivative” meansany pharmaceutically acceptable salt, ester, or salt of such ester, of acompound of this invention or any other compound which, uponadministration to a recipient, is capable of providing (directly orindirectly) a compound of this invention or an anti-virally activemetabolite or residue thereof.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of suitable acids includehydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric,maleic, phosphoric, glycollic, lactic, salicylic, succinic,toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic,benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acids.Other acids, such as oxalic, while not in themselves pharmaceuticallyacceptable, may be employed in the preparation of salts useful asintermediates in obtaining the compounds of the invention and theirpharmaceutically acceptable acid addition salts.

Salts derived from appropriate bases include alkali metal (e.g.,sodium), alkaline earth metal (e.g., magnesium), ammonium and N—(C₁₋₄alkyl)₄ ⁺ salts.

The term “thiocarbamates” refers to compounds containing the functionalgroup N—SO₂—O.

The compounds of this invention contain one or more asymmetric carbonatoms and thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers. Allsuch isomeric forms of these compounds are expressly included in thepresent invention. Each stereogenic carbon may be of the R or Sconfiguration. The explicitly shown hydroxyl is also preferred to be synto D, in the extended zig-zag conformation between the nitrogens shownin compounds of formula I.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and administration to a mammalby methods known in the art. Typically, such compounds are stable at atemperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

The compounds of the present invention may be used in the form of saltsderived from inorganic or organic acids. Included among such acid salts,for example, are the following: acetate, adipate, alginate, aspartate,benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate,pectinate, persulfate, 3-phenylpropionate, picrate, pivalate,propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate.

This invention also envisions the quaternization of any basicnitrogen-containing groups of the compounds disclosed herein. The basicnitrogen can be quaternized with any agents known to those of ordinaryskill in the art including, for example, lower alkyl halides, such asmethyl, ethyl, propyl and butyl chloride, bromides and iodides; dialkylsulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; longchain halides such as decyl, lauryl, myristyl and stearyl chlorides,bromides and iodides; and aralkyl halides including benzyl and phenethylbromides. Water or oil-soluble or dispersible products may be obtainedby such quaternization.

The novel sulfonamides of this invention are those of formula I:

wherein:

A is selected from the group consisting of H; Het; —R¹-Het; —R¹—C₁-C₆alkyl, which may be optionally substituted with one or more groupsselected from the group consisting of hydroxy, C₁-C₄ alkoxy, Het,—O-Het, —NR²—CO—N(R²)(R²) and —CO—N(R²) (R²); and —R¹—C₂-C₆ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het, —O-Het,—NR²—CO—N(R²)(R²) and —CO—N(R²)(R²);

each R¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)— and—NR²—C(O)—C(O)—;

each Het is independently selected from the group consisting of C₃-C₇cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturatedor unsaturated heterocycle, containing one or more heteroatoms selectedfrom N, N(R²), O, S and S(O)_(n), wherein said heterocycle mayoptionally be benzofused; and wherein any member of said Het may beoptionally substituted with one or more substituents selected from thegroup consisting of oxo, —OR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)(R²), —S(O)₂—N(R²)(R²), —N(R²)—C(O)—R², —C(O)—R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—Ar, methylenedioxy, —N(R²)—S(O)₂(R²),halo, —CF₃, —NO₂, Ar and —O—Ar;

each R² is independently selected from the group consisting of H andC₁-C₃ alkyl optionally substituted with Ar;

B, when present, is —N(R²)—C(R³)(R³)—C(O)—;

x is 0 or 1;

each R³ is independently selected from the group consisting of H, Het,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl,wherein any member of said R³, except H, may be optionally substitutedwith one or more substituents selected from the group consisting of—OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Het, —CN, —SR², —C₂R²,NR²—C(O)—R²;

each n is independently 1 or 2;

D and D′ are independently selected from the group consisting of Ar;C₁-C₄ alkyl, which may be optionally substituted with one or more groupsselected from C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₂-C₄ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar;C₃-C₆ cycloalkyl, which may be optionally substituted with or fused withAr; and C₅-C₆ cycloalkenyl, which may be optionally substituted with orfused with Ar;

each Ar is independently selected from the group consisting of phenyl;3-6 membered carbocyclic ring and 5-6 membered heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) andN(R²), wherein said carbocyclic or heterocyclic ring may be saturated orunsaturated and optionally substituted with one or more groups selectedfrom the group consisting of oxo, —OR², —R², —N(R²)(R²), —N(R²)—C(O)—R²,—R²—OH, —CN, —CO₂R², —C(O)—N(R²)(R²), halo and —CF₃;

E is selected from the group consisting of Het; O-Het; Het-Het; —O—R³;—NR²R³; C₁-C₆ alkyl, which may be optionally substituted with one ormore groups selected from the group consisting of R⁴ and Het; C₂-C₆alkenyl, which may be optionally substituted with one or more groupsselected from the group consisting of R⁴ and Het; C₃-C₆ saturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and C₅-C₆ unsaturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and

each R⁴ is independently selected from the group consisting of —OR²,—C(O)—NHR², —S(O)₂—NHR², halo, —NR²—C(O)—R² and —CN.

Except where expressly provided to the contrary, as used herein, thedefinitions of variables A, R¹-R⁴, Het, B, x, n, D, D′, Ar and E are tobe taken as they are defined above for the compounds of formula I.

According to one embodiment of this invention, a subclass of compoundsare those compounds of formula I, and pharmaceutically acceptable saltsthereof, wherein:

A is selected from the group consisting of H; —R¹-Het; —R¹—C₁-C₆ alkyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het and —O-Het; and—R¹—C₂-C₆ alkenyl, which may be optionally substituted with one or moregroups selected from hydroxy, C₁-C₄ alkoxy, Het and —O-Het;

each R¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—CO—, —O—S(O)₂— and —NR²—S(O)₂—;

each Het is independently selected from the group consisting of C₃-C₇cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturatedor unsaturated heterocycle, containing one or more heteroatoms selectedfrom N, O and S, which may optionally be benzofused; wherein any memberof said Het may be optionally substituted with one or more substituentsselected from the group consisting of oxo, —OR², —R², —N(R²)₂, —R²—OH,—CN, —CO₂R², —C(O)—N(R²)₂ and —S(O)₂—N(R²)₂;

each R is independently selected from the group consisting of H andC₁-C₃ alkyl;

B, when present, is —NH—CH(R³)—C(O)—;

x is 0 or 1;

R³ is selected from the group consisting of Het, C₁-C₆ alkyl, C₂-C₆alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl, wherein any member ofsaid R³ may be optionally substituted with one or more substituentsselected from the group consisting of —OR², —C(O)—NH—R²,—S(O)_(n)—N(R²)₂, Het and —CN;

n is 1 or 2;

D and D′ are independently selected from the group consisting of Ar;C₁-C₄ alkyl, which may be optionally substituted with C₃-C₆ cycloalkylor Ar; C₂-C₄ alkenyl, which may be optionally substituted with C₃-C₆cycloalkyl or Ar; C₃-C₆ cycloalkyl, which may be optionally substitutedor fused with Ar; and C₅-C₆ cycloalkenyl, which may be optionallysubstituted or fused with Ar; with the proviso that when D is attachedto N, D may not be methyl or C₂ alkenyl;

Ar is selected from the group consisting of phenyl; 3-6 memberedcarbocyclic ring and 5-6 membered heterocyclic ring containing one ormore heteroatoms selected from O, N and S, wherein said carbocyclic orheterocyclic ring may be saturated or unsaturated and optionallysubstituted with one or more groups selected from the group consistingof oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)R², —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)₂, halo and —CF₃;

E is selected from the group consisting of Het; —O—R³; —NR²R⁵; C₁-C₆alkyl, which may be optionally substituted with one or more R⁴ or Het;C₂-C₆ alkenyl, which may be optionally substituted with one or more R⁴or Het; C₃-C₆ saturated carbocycle, which may optionally be substitutedwith one or more R⁴ or Het; and C₅-C₆ unsaturated carbocycle, which mayoptionally be substituted with one or more R⁴ or Het;

each R⁴ is independently selected from the group consisting of —OR²,—C(O)—NHR², —S(O)₂—NHR², halo and —CN; and

each R⁵ is independently selected from the group consisting of H and R³,with the proviso that at least one R⁵ is not H.

A preferred subclass of compounds of this invention are those compoundsof formula I having a molecular weight of less than about 700 g/mole.More preferably, the subclass of compounds of formula I have a molecularweight of less than about 600 g/mole.

Other preferred subclasses of this invention are those compounds offormulas XXII, XXIII and XXXI:

wherein A, R³, Het, D, D′, x and E are as defined above for compounds offormula I. For ease of reference, the two R³ moieties present in formulaXXXI have been labeled R³ and R³′.

For compounds of formula XXII, most preferred compounds are thosewherein A is R¹-Het and D′ is C₁-C₃ alkyl or C₃ alkenyl, wherein saidalkyl or alkenyl may optionally be substituted with one or more groupsselected from the group consisting of C₃-C₆ cycloalkyl, —OR², —O—Ar andAr (with all other variables being defined as above for compounds offormula I). For compounds of formula XXIII, most preferred compounds arethose wherein R³ is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₅-C₆ cycloalkyl, C₅-C₆cycloalkenyl or a 5-6 membered saturated or unsaturated heterocycle,wherein any member of said R³ may be optionally substituted with one ormore substituents selected from the group consisting of —OR²,—C(O)—NH—R², —S(O)_(n)N(R²)(R²), Het, —CN, —SR², —C(O)₂R² andNR²—C(O)—R² and D′ is C₁-C₃ alkyl or C₃ alkenyl, wherein said alkyl oralkenyl may optionally be substituted with one or more groups selectedfrom the group consisting of C₃-C₆ cycloalkyl, —OR², —O—Ar and Ar (withall other variables being defined as above for compounds of formula I).

For compounds of formula XXXI, most preferred compounds are thosewherein A is R¹-Het, each R³ is independently C₁-C₆ alkyl which may beoptionally substituted with a substituent selected from the groupconsisting of —OR², —C(O)—NH—R², —S(O)_(n)N(R²) (R²), Het, —CN, —SR²,—CO₂R² and —NR²—C(O)—R²; D′ is C₁-C₄ alkyl, which may be optionallysubstituted with a group selected from the group consisting of C₃-C₆cycloalkyl, —OR², —O—Ar; and E is Het, Het-Het and —NR²R³.

Sulfonamides of this invention include the following specific compoundscontained in Tables I-VI. In Tables I-IV and VI, A is attached throughthe right-most bond, unless otherwise expressly noted. All othersubstituents in Tables I-VI are attached via the left-most bond, unlessotherwise expressly noted.

TABLE I

COMPOUND A R³ D′ E 1

2

3

4

5

6

7

8

CH₃ 9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

TABLE II

COMPOUND A D′ E 27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

CH₃

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

CH₃

127

CH₃

128

CH₃

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

1001

1002

1003

1004

1005

—CH₃

1006

1007

1008

1009

1010

1011

1012

1013

1014

1015

TABLE III

COMPOUND A D′ E 186

187

188

TABLE IV

COMPOUND A 189

190

TABLE V

COM- POUND E 191

192

193

194

TABLE VI

COMPOUND D D′ 195

196

Preferred compounds of this invention are*:

-   (S)-N-1-(3-((3-Acetylamino-4-fluoro-benzenesulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    and    (S)-N-1-(3-((4-Acetylamino-3-fluoro-benzenesulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compounds 2);-   (S)-N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 5);-   (S)-N-1-(1-Benzyl-3-(benzyl-(5-isoxazol-3-yl-thiophene-2-sulfonyl)-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 6);-   (S)-N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 9);-   N-1-(1-(S)-Benzyl-3-(benzyl-(3-sulfamoyl-benzenesulfonyl)-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 10);-   (S)-N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobutyl-(5-pyridin-2-yl-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 12);-   (S)-N-1-(3-((4-Benzenesulfonyl-thiophene-2-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 13);-   (S)-N-1-(1-(S)-Benzyl-3-((4-fluoro-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 14);-   (S)-N-1-(3-((4-Acetylamino-3-fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 15);-   (S)-N-1-(3-((3-Acetylamino-4-fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 16);-   (S)-N-1-(1-(S)-Benzyl-3-((4-acetylamino-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 17);-   (S)-N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 18);-   (S)-N-1-(3-((3-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 19);-   (S)-N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 20);-   N-1-((1S-2    syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfonylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 21);-   N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinamide    (compound 22);-   N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-4-yl-methoxycarbonyl)-succinamide    (compound 23);-   N-1-(3-((4-Fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinamide    (compound 26);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 35);-   3,4-Dichloro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 37);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 44);-   2,4-Dimethyl-thiazole-5-sulfonic    acid-(1,1-dimethyl-ethoxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-amide (compound 46);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 48);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    and 4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compounds 52);-   Benzo(1,2,5)oxadiazole-5-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-isobutylamide    (compound 66);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl-phenyl)-acetamide    and N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compounds 86);-   N-(2-Fluoro-5-(((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 88);-   N-(3-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 91);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 93);-   N-(4-(((syn)-2-Hydroxy-(S)-4-phenyl-3-((tetrahydro-furan-(R)-3-yl)-oxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 94);-   4-Fluoro-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(R)-3-ylmethoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    and 4-Fluoro-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(S)-3-ylmethoxycarbonylamino)-butyl)-N-isobutyl    -benzenesulfonamide (compounds 97);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 98);-   4-Chloro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-benzenesulfonamide    (compound 99);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide    (compound 100);-   4-Fluoro-N-(2-(syn)-hydroxy-3-((2-oxazolidon-(S)-4-yl)-methoxycarbonylamino)-4-(S)-phenyl-butyl)-N-isobutyl-benzenesulfonamide    (compound 109);-   Benzene-1,3-disulfonic acid 1-amide 3-((2    syn,3S)-2-hydroxy-4-phenyl-3-(3-(S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 112);-   Furan-3-sulfonic acid (2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 113);-   N-((3-Allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-N-cyclopentylmethyl-4-fluoro-benzenesulfonamide    (compound 114);-   N-Cyclopentylmethyl-N-((3-ethoxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-4-fluoro-benzenesulfonamide    (compound 115);-   4-Chloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 116);-   4-Chloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3yl-methoxycarbonyl)-butyl)-benzenesulfonamide    (compound 118);-   N-(4-(Cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide    (compound 125);-   3-Chloro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 138);-   4-Chloro-N-cyclopentylmethyl-N-(2-(syn)-hydroxy-3-((2-oxazolidon-4-(S)-yl-methyl)-oxycarbonylamino)-4-phenyl-butyl)-benzenesulfonamide    (compound 139);-   N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 140);-   N-((3-allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide    (compound 141);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(3-pyridin-3-yl-methoxycarbonylamino)-butyl-4-methoxy-benzenesulfonamide    (compound 142);-   Pyridine-3-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide,    trifluoroacetic acid salt (compound 144);-   5-Isoxazol-3-yl-thiophene-2-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 145);-   N-(4-((3-(Allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-cyclopentylmethylsulfamoyl)-phenyl)-acetamide    (compound 146);-   N-(4-(Cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide    (compound 147);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 148);-   Pyridine-3-sulfonic acid cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-amide    (compound 149);-   Piperidine-1-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 150);-   N-4-((2-(syn)-Hydroxy-3-((2-methoxymethyl-allyloxycarbonylamino)-4-(S)-phenyl-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 155);-   1-Acetyl-2,3-dihydro-1H-indole-6-sulfonic acid    ((allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-cyclopentylmethyl-amide (compound    156);-   1-Acetyl-2,3-dihydro-1H-indole-6-sulfonic acid cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-amide    (compound 157);-   N-Cyclohexylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 158);-   N-Cyclohexylmethyl-4-fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 159);-   N-(4-(Cyclohexylmethyl)-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-acetamide    (compound 160);-   N-((2 syn,3S)-2-Hydroxy-4-phenyl-3-(pyridin-4-yl    -methoxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide    (compound 163);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((syn)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methyl-benzenesulfonamide    (compound 165);-   N-cyclopentylmethyl-4-hydroxy-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-benzenesulfonamide    (compound 166);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-nitro-benzenesulfonamide    (compound 167);-   4-Amino-N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 168);-   N-Cyclopentylmethyl-4-hydroxy-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 169);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-nitro-benezensulfonamide    (compound 170);-   4-Amino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 171);-   2,4-Diamino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 173);-   4-Hydroxy-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 175);-   N-Cyclopentylmethyl-4-fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 182);-   3,4-Dichloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 183);-   Benzyloxycarbonyl-(L)-isoleucine-N-(5-((3-amino-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-sulfamoyl)-2-fluoro-phenyl)-acetamide    (compound 187);-   N-((2    syn,3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide    (compound 195);-   and compounds 1001 through 1015. * As can be appreciated by those of    ordinary skill in the art, different conventions are used in naming    chemical compounds. Because of possible discrepencies in the art of    chemical nomenclature, the structyres show in Tables I-VI hererin    are controlling for the definition of compounds 1-195 and 1001-1015    of this invention.

More preferred compounds of this invention are:

-   (S)-N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobutyl-(5-pyridin-2-yl-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 12);-   (S)-N-1-(1-(S)-Benzyl-3-((4-fluoro-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 14);-   (S)-N-1-(3-((4-Acetylamino-3-fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 15);-   (S)-N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 20);-   N-1-((1S-2    syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfonylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 21);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 48);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide    (compound 100);-   4-Chloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 116);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 140);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(3-pyridin-3-yl-methoxycarbonylamino)-butyl-4-methoxy-benzenesulfonamide    (compound 142);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 148);-   N-Cyclohexylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 158);-   N-(4-(Cyclohexylmethyl)-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-acetamide    (compound 160);-   N-cyclopentylmethyl-4-hydroxy-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-benzenesulfonamide    (compound 166);-   4-Amino-N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 168);-   4-Amino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 171);-   2,4-Diamino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 173);-   4-Hydroxy-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 175); and-   N-((2    syn,3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide    (compound 195).

The sulfonamides of this invention may be synthesized using conventionaltechniques. Advantageously, these compounds are conveniently synthesizedfrom readily available starting materials.

The compounds of this invention are among the most readily synthesizedHIV protease inhibitors known. Previously described HIV proteaseinhibitors often contain four or more chiral centers, numerous peptidelinkages and/or require air-sensitive reagents (such as organometalliccomplexes) to effect their synthesis. The relative ease with which thecompounds of this invention can be synthesized represents an enormousadvantage in the large scale production of these compounds.

In general, sulfonamides of formula I are conveniently obtained fromα-amino acid derivatives having the general formulaA-(B)_(x)-NH—CH(D)-COOH, wherein A, B, X and D are defined as above forthe compounds of formula I. Such α-amino acid derivatives are oftencommercially available or may be conveniently prepared from commerciallyavailable α-amino acid derivatives using known techniques. See, forexample, T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis”, 2nd Ed., John Wiley and Sons (1991). Although this inventionenvisions the use of racemic mixtures of such starting materials, whenx=0, a single enantiomer in the S configuration is preferred.

Using known techniques, the α-amino acid derivative of general formulaA-(B)_(x)-NH—CH(D)-COOH may be readily converted to an amino ketonederivative of general formula A-(B)_(x)-NH—CH(D)-CO—CH₂—X, wherein X isa leaving group which suitably activates the α-carbon (i.e., makes themethylene susceptible to nucleophilic attack). Suitable leaving groupsare well known in the art and include halides and sulfonates, such asmethanesulfonate, trifluoromethanesulfonate or 4-toluenesulfonate X mayalso be a hydroxyl which is converted in situ to a leaving group (e.g.by treatment with a trialkyl- or triarylphosphine in the presence of adialkylazodicarboxylate). Methods for the formation of such amino ketonederivatives also are well known to those of skill in the art (see, forexample, S. J. Fittkau, J. Prakt. Chem., 315, p. 1037 (1973)).Alternatively, certain amino ketone derivatives are commerciallyavailable (e.g., from Bachem Biosciences, Inc., Philadelphia, Pa.).

The amino ketone derivative may then be reduced to the correspondingamino alcohol, represented by the formulaA-(B)_(x)-NH—CH(D)-CH(OH)—CH₂—X. Many techniques for reduction of aminoketone derivatives such as A-(B)_(x)-NH—CH(D)-CO—CH₂—X are well known tothose of ordinary skill in the art (Larock, R. C. “Comprehensive OrganicTransformations”, pp. 527-547, VCH Publishers, Inc.© 1989 and referencescited therein). A preferred reducing agent is sodium borohydride. Thereduction reaction is conducted at a temperature of from about −40° C.to about 40° C. (preferably, at about 0° C. to about 20° C.), in asuitable solvent system such as, for example, aqueous or neattetrahydrofuran or a lower alcohol, such as methanol or ethanol.Although this invention envisions both stereospecific andnon-stereospecific reduction of the amino ketone derivativeA-(B)_(x)-NH—CH(D)-CO—CH₂—X, stereoselective reduction is preferred.Stereoselective reduction may be accomplished by use of chiral reagentsknown in the art. In the present invention, stereoselective reductionmay be conveniently achieved, for instance, under non-chelating reducingconditions, where chiral induction of the newly formed hydroxyl group isset by the stereochemistry of the D group (i.e., Felkin-Ahn addition ofhydride). We particularly prefer stereoselective reductions wherein theresulting hydroxyl is syn to D. We have found that when the hydroxylgroup is syn to D, the final sulfonamide product is an HIV proteaseinhibitor of higher potency than the anti diastereomer.

The hydroxyl group of the amino alcohol may optionally be protected byany known oxygen protecting group (such as trialkylsilyl, benzyl, oralkyloxymethyl) to yield a protected amino alcohol having the formulaA-(B)_(x)-NH—CH(D)-C(OR⁶)—CH₂—X, wherein R⁶ is H or any suitable hydroxyprotecting group. Several useful protecting groups are described in T.W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2dEd., John Wiley and Sons (1991).

The amino alcohol may then be reacted with a nucleophilic amine compoundto form an intermediate of formula III:

wherein D and R⁶ are as described above, and L is either D′ (asdescribed for compounds of formula I) or hydrogen.

In a particularly advantageous synthetic scheme, simultaneous activationof the methylene and protection of the alcohol may be accomplished byforming an N-protected amino epoxide from the oxygen and its adjacentmethylene to give an intermediate of formula II:

wherein A, B and D are as defined above for compounds of formula I.Suitable solvent systems for preparing the N-protected amino epoxideinclude ethanol, methanol, isopropanol, tetrahydrofuran, dioxane,dimethyl formamide and the like (including mixtures thereof). Suitablebases for producing the epoxide include alkali metal hydroxides,potassium t-butoxide, DBU and the like. A preferred base is potassiumhydroxide.

Reaction of the N-protected amino epoxide or other suitably activatedintermediates with an amine is carried out neat, i.e. in the absence ofsolvent, or in the presence of a polar solvent such as lower alkanols,water, dimethylformamide or dimethylsulfoxide. The reaction can becarried out conveniently between about 0° C. and 120° C., preferablybetween about 20° C. and 100° C. Alternatively, the reaction may becarried out in the presence of an activating agent, such as activatedalumina in an inert solvent, preferably an ether, such as diethyl ether,tetrahydrofuran, dioxane, or tert-butyl methyl ether, conveniently fromabout room temperature to about 110° C., as described by Posner andRogers, J. Am. Chem. Soc., 99, p. 8208 (1977). Other activating reagentsinclude lower trialkylaluminum species, such as triethylaluminum, ordialkylaluminum halide species, such as diethylaluminum chloride(Overman and Flippin, Tetrahedron Letters, p. 195 (1981)). Reactionsinvolving these species are conveniently carried out in inert solventssuch as dichloromethane, 1,2-dichloroethane, toluene, or acetonitrilebetween about 0° C. and about 110° C. Further methods of displacingleaving groups, or opening epoxides with amines or their equivalentssuch as azides or timethylsilyl cyanide (Gassman and Guggenheim, J. Am.Chem. Soc. 104, p. 5849 (1982)), are known and will be apparent to thoseof ordinary skill in the art.

Compounds of formulae II and III, and functionality-protectedderivatives thereof, are useful as intermediates for the preparation ofcompounds of formula I. In those cases where L represents D′, compoundsof formula III may be converted to compounds of formula I by reactionwith sulfonyl-activated species to form sulfonamides, sulfonyl ureas,thiocarbamates and the like. Methods for preparing suchsulfonyl-activated species are well within the ordinary skill of theart. Typically, sulfonyl halides are used to obtain sulfonamides. Manysulfonyl halides are commercially available; others may be easilyobtained using conventional synthetic techniques (Gilbert, E. E. “RecentDevelopments in Preparative Sulfonation and Sulfation” Synthesis 1969: 3(1969) and references cited therein; Hoffman, R. V.“M-Trifluoromethylbenzenesulfonyl Chloride” Org. Synth. Coll. Vol. VII,John Wiley and Sons (1990); Hartman, G. D. et. al. “4-SubstitutedThiophene-and Furan-2-sulfonamides as Topical Carbonic AnhydraseInhibitors” J. Med. Chem., 35, p. 3822 (1992) and references citedtherein. Sulfonyl ureas are usually obtained by the reaction of an aminewith sulfuryl chloride or a suitable equivalent such assulfuryl-bis-imidazole or sulfuryl-bis-N-methyl imidazole.Thiocarbamates are typically obtained by the reaction of an alcohol withsulfuryl chloride or a suitable equivalent such assulfuryl-bis-imidazole or sulfuryl-bis-N-methyl imidazole.

In the case of compounds of formula III wherein L is hydrogen,conversion of the resultant primary amine to a secondary amine may becarried out by known techniques. Such techniques include reaction withan alkyl halide of alkyl sulfonate, or by reductive alkylation with analdehyde or carboxylic acid or activated derivative thereof using, forinstance, catalytic hydrogenation or sodium cyanoborohydride (Borch etal., J. Am. Chem. Soc., 93, p. 2897 (1971)). Alternatively, the primaryamine may be acylated followed by reduction with borane or anothersuitable reducing reagent, for example, as described by Cushman et al.,J. Org. Chem., 56, p. 4161 (1991). This technique is especially usefulin compounds of formula III where B is absent and A represents aprotecting group such as tert-butoxycarbonyl (Boc) or benzyloxycarbonyl(Cbz).

If variable A of a particular compound of formula I represents aremovable protecting group, removal of that group followed by reactionof the resulting amine with an appropriate activated reagent willadvantageously yield a different compound of formula I. For instance,reaction with an activated carboxylate, such as an acyl halide (e.g.,acid fluorides, acid chlorides, and acid bromides), an activated estersuch as nitrophenyl ester or 1-hydroxysuccinimide (HOSu) ester, ananhydride such as the symmetrical anhydride or isobutyl anhydride, ormixed carbonic-phosphoric or carbonic-phosphinic anhydrides, will yieldthe corresponding amide. Ureas may be obtained by reaction withisocyanates or amines in the presence of bis-activated carbonic acidderivatives such as phosgene or carbonyldiimdazole. Carbamates may beobtained by reaction with chlorocarbonates, with carbonates esterifiedwith leaving groups such as 1-hydroxybenzotriazole (HOBT) or HOSu, orwith alcohols in the presence of bis-activated carbonic acid derivativessuch as phosgene or carbonyldiimdazole. It will be readily recognizedthat in order to facilitate specific reactions, the protection of one ormore potentially reactive groups followed by subsequent removal of thatgroup may be required. Such modification to the reaction schemesoutlined above are within the ordinary skill of the art.

If variable B of a particular compound of formula I is absent andvariable A of that compound represents a removable protecting group,removal of A, followed by reaction of the resulting amine with an aminoacid or suitably N-protected derivative thereof, followed by asubsequent reaction of the free α-amine if present, as described above,will yield a further compound of formula I. The addition of amino acidsand their derivatives is accomplished by well known methods of peptidesynthesis. Some of these methods are generally set forth in Bodanszkyand Bodanszky, “The Practice of Peptide Synthesis”, Springer-Verlag,Berlin, Germany (1984) and in the “The Peptides”, Gross and Meinhofer(Eds); Academic Press, 1979, Vols. I-III, which are incorporated hereinby reference.

Typically, for solution phase synthesis of peptides, the α-amine of theamino acid to be coupled is protected by Boc, Cbz, allyloxycarbonyl(Alloc) or 9-fluorenylmethoxycarbonyl (Fmoc), while the free carboxyl isactivated by reaction with a carbodiimide such asdicyclohexylcarbodiimide (DCC),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), ordiisopropylcarbodiimide (DIC), optionally in the presence of a catalystsuch as HOBT, HOSu, or dimethylaminopyridine (DMAP). Other methods whichproceed through the intermediacy of activated esters, acid halides,enzyme-activated amino acids and anhydrides includingN-carboxy-anhydrides, symmetrical anhydrides, mixed carbonic anhydrides,carbonic-phosphinic and carbonic-phosphoric anhydrides are alsosuitable. After the peptide has been formed, protecting groups may beremoved by methods described in the references listed above, such as byhydro-genation in the presence of a palladium, platinum or rhodiumcatalyst, treatment with sodium in liquid ammonia, hydrochloric,hydrofluoric, hydrobromic, formic, trifluoromethanesulfonic, ortrifluoroacetic acid, secondary amines, fluoride ion, trimethylsilylhalides including bromide and iodide, or alkali.

One particularly useful synthetic scheme for producing sulfonamides offormula XV is shown below:

Compounds of formula X may be advantageously synthesized from readilyavailable starting materials (see D. P. Getman, J. Med. Chem., 36, p.288 (1993)). Each step of the above synthetic scheme may be carried outas generally described above.

A particularly useful synthetic scheme for producing the preferredsulfonamides of formula XXII is shown below:

Compounds of formula XX may be advantageously synthesized from readilyavailable starting materials (see B. E. Evans et al., J. Org. Chem., 50,p. 4615 (1985)). Each step of the above synthetic scheme may be carriedout as generally described above.

After converting a compound of formula XX to a compound of formula XXI,as detailed in the previous reaction scheme, the compound of formula XXImay alternatively be reacted with an amino acid or amino acidderivative, as described generally above, to yield a preferred compoundof formula XXXI. A particularly useful synthetic scheme utilizing thisstrategy is set forth below:

As can be appreciated by the skilled artisan, the above syntheticschemes are not intended to comprise a comprehensive list of all meansby which the compounds described and claimed in this application may besynthesized. Further methods will be evident to those of ordinary skillin the art.

The compounds of this invention may be modified by appending appropriatefunctionalites to enhance selective biological properties. Suchmodifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds of formula I are characterized by a superior ability toinhibit HIV protease activity and viral replication. We believe thatthis is due to specific steric and electronic interactions between theprotease and compounds of formula I. This belief stems from our analysisof the structural basis for the activity of compounds of formula I, inview of the known crystal structures of HIV protease and boundinhibitors, such as the structure reported in Miller et al. “Structureof Complex of Synthetic HIV-1 Protease with a Substrate-Based Inhibitorat 2.3 Å Resolution”, Science, vol. 246, pp. 1149-1152 (1989), which isincorporated herein by reference, as well as structures determined inour laboratories. According to these structures, the active site of HIVaspartyl protease is defined by a deep groove containing subpockets foraccommodation of various side chains of the protease substrate—referredto as P₁-P_(n) and P₁′-P_(n)′, according to conventional proteasenomenclature. In the center of the groove, lie two aspartic acidresidues (Asp25 and Asp25′ according to the numbering system of Milleret al.) in a manner typical of the active site aspartates of knownaspartyl proteases, which are believed to be the catalytic residues ofthe enzyme. The groove is covered by two C₂-symmetrically disposed“flaps” which also make various direct and indirect contacts with boundsubstrates.

We believe that the substituents A, D, D′ and E of the compounds offormula I associate with HIV protease by way of hydrophobic forces inthe binding pockets of the enzyme. We also believe that the sulfonamidegroup hydrogen binds tightly to a water molecule held by hydrogen bondsto the flaps of the protease (“the flap water molecule”; water molecule511, according to the Miller et al. numbering system).

In view of the above discovery, an alternative embodiment of thisinvention relates to novel HIV protease inhibitors possessing certainstructural and physicochemical features. We have discovered thatcompounds possessing the following novel combination of features aresurprisingly effective HIV protease inhibitors:

(1) a first and a second hydrogen bond acceptor moiety, at least one ofwhich is more highly polarizable than a carbonyl, said moieties beingthe same or different, and being capable of hydrogen bonding with thehydrogen atoms of the flap water molecule of an HIV aspartyl proteasewhen the compound is bound thereto;

(2) substantially hydrophobic moieties which associate with the P₁ andP₁′ binding pockets of said HIV aspartyl protease when the compound isbound thereto;

(3) a third hydrogen bonding moiety, which may be either a hydrogen bonddonor or acceptor, capable of simultaneously hydrogen bonding to Asp25and Asp25′ of said HIV aspartyl protease when the compound is boundthereto;

(4) an additional occupied volume of space of at least 100 Å³ when thecompound is bound to the active site of said HIV aspartyl protease, saidspace overlapping with the volume of space that would be filled by anative substrate of said HIV aspartyl protease or a nonhyrolyzableisostere thereof;

(5) a deformation energy of binding of the compound to said HIV aspartylprotease of not greater than 10 kcal/mole; and

(6) a neutral or favorable enthalpic contribution from the sum of allelectrostatic interactions between the compound and the protease whenthe compound is bound to said HIV aspartyl protease.

Compounds having the above-cited features can be readily identified ordesigned by one of ordinary skill in the art using a combination ofchemical reasoning and computational methods. For example, those ofordinary skill in the art can readily identify or choose hydrogenbonding and hydrophobic moieties or groups required in features (1)-(3),while features (4)-(6) can be ascertained using well known computationalmethods for determination of structural (e.g. conformational) andenergetic properties of molecules.

Furthermore, compounds characterized by features (1) through (6) listedabove may be obtained using any conventional technique, includingchemical synthesis and natural product isolation. We prefer using thesynthetic schemes detailed above for compounds of formula I.

We have discovered that when an HIV protease inhibitor forms hydrogenbonds to the flap water molecule through two hydrogen bonding moieties,at least one of which is more highly polarizable than a carbonyl, theability of those compounds to inhibit HIV protease activity isdramatically improved, as compared with conventional HIV proteaseinhibitors.

While not wishing to be bound by theory, we believe that the stronghydrogen bonds that form between the flap water molecule and the twohydrogen bonding moieties, at least one of which is more highlypolarizable than a carbonyl, lower the overall binding energy of theinhibitor. Most HIV protease inhibitors known in the art utilize onlycarbonyl groups for hydrogen bonding to the flap water molecule and are,thus, inferior to those of the present invention. We believe that theincreased polarization that results from the large dipole moment of thehighly polarizable hydrogen bonding moiety (as compared to the dipolemoment of a carbonyl moiety) creates a stronger and tighter hydrogenbond with the flap water molecule. We prefer to utilize tetravalentoxygenated sulfur, hexavalent oxygenated sulfur and pentavalentoxygenated phosphorus as the highly polarizable hydrogen bonding moiety.Tetravalent oxygenated sulfur and hexavalent oxygenated sulfur are morepreferred as the highly polarizable hydrogen bonding moiety. Hexavalentoxygenated sulfur (—SO₂—) is most preferred.

We have found that when the highly polarizable hydrogen bonding moietyis a sulfonamide, the overall binding energy of the inhibitor isparticularly low. We believe that this increased stability is due toparticular conformational characteristics of the sulfonamide S—N bond.Specifically, the sulfonamide S—N bond exists in only two low-energyrotamers (see J. B. Nicholas et al., J. Phys. Chem., 95, p. 9803 (1991)and R. D. Bindal et al., J. Am. Chem. Soc., 112, p. 7861 (1990)). Thishas the effect of locking that portion of the molecule into a favorableconformation wherein one or both of the highly polarized S═O oxygens canbe involved in hydrogen bonding interactions with the flap water.

The remaining five structural and physicochemical features recited above(i.e., features (2) through (6)) are generally recognized in the art toimprove the ability of a compound to competitively inhibit HIV proteaseactivity. Although there are several other features thought to increasethe inhibitory property (such as binding of the inhibitor backbone tothe enzyme), we have discovered that the combination of the fiveabove-cited elements alone, together with novel element (1), typifieseffective HIV protease inhibitors.

In general, the binding energy of a particular protease inhibitor islowered when hydrophobic moieties on the inhibitor are located so as toassociate with the enzyme's hydrophobic binding pockets. In the case ofHIV-1 protease, the location and nature of the P₁ and P₁′ bindingpockets are known to those of ordinary skill in the art (see, forexample, M. Miller et al., cited above). Substantially hydrophobic sidechains which fit into the well defined P₁ and P₁′ binding pockets arealso known to those in the art. Preferred side chains are located within4 Å of the enzyme when bound to HIV protease. Preferred hydrophobic sidechains include those substantially similar to those of hydrophobicnatural and unnatural α-amino acids, including alanine, valine, leucine,isoleucine, methionine, phenylalanine, α-amino isobutyric acid,alloisoleucine, tyrosine, and tryptophan. Insofar as a portion of thisside chain is in contact with bulk solvent or protrudes out of theenzyme, it is not considered to be wholly within P₁ or P₁′ and maycontain polar functionality such as a charged amine at that location.

It has also been established in the art that the presence of a hydroxylgroup within hydrogen bond proximity to the two catalytic aspartic acidresidues of HIV protease (Asp25 and Asp25′) is an important feature ofan effective HIV protease inhibitor (see, for example, R. Bone et al.,“X-ray Crystal Structure of the HIV Protease Complex with L-700, 417, anInhibitor with Pseudo C₂ Symmetry”, J. Am. Chem. Soc., 113, pp. 9382-84(1991)). It is further understood that the geometry of the Asp-bindinghydrogen bonding moiety is of particular importance. Although we preferto use a hydroxyl group at this position, any hydrogen bonding moietythat is capable of forming hydrogen bonds with the Asp residues isacceptable. Such hydrogen bonding moieties are known to those of skillin the art (e.g., phosphinic acid (D. Grobelny et al., Biochem. Biophys.Res. Commun., 169, p. 1111 (1990)).

It is further understood that binding of competitive inhibitors to HIVprotease is optimally accomplished by having the inhibitor traverse avolume overlapping that occupied by the native polypeptide substratewhen it is bound to the active site of the enzyme. Effective HIVprotease inhibitors typically have a relatively small difference inenergy between their bound and free states (i.e., a small deformationenergy of binding). The most preferred HIV protease inhibitors of thisinvention have a deformation energy of binding of not greater than 10kcal/mole (preferably, not greater than 7 kcal/mole). It should benoted, however, that HIV protease inhibitors may interact with HIVprotease in more than one conformation which is similar in overallbinding energy (see K. H. M. Murthy, J. Biol. Chem., 267, (1992)). Inthose cases, the deformation energy of binding is taken to be thedifference between the energy of the free compound and the averageenergy of the conformations observed when the inhibitor binds to theenzyme.

Furthermore, it is understood that the most effective proteaseinhibitors also lack repulsive electrostatic interaction with the targetprotease in their bound state. Such non-complementary (e.g.,electrostatic) interactions include repulsive charge-charge,dipole-dipole and charge-dipole interactions. Specifically, in the mostpreferred HIV protease inhibitors of this invention, the sum of allelectrostatic interactions between the compound and the enzyme when thecompound is bound to HIV protease makes a neutral or favorablecontribution to the enthalpy of binding.

Preferred compounds characterized by the above features (1)-(6) arecompounds of formula XL:Z¹-Q¹-L¹-M-L²-Q²-Z²  (XL)wherein:

Q¹ and Q² are independently hydrogen bond acceptor moieties capable ofbinding with the hydrogen atoms of the flap water molecule of an HIVaspartyl protease, with the proviso that at least one of Q¹ or Q² ismore highly polarizable than a carbonyl;

M is a hydrogen bonding moiety, which may be either a hydrogen bonddonor or acceptor, capable of simultaneously hydrogen bonding to Asp25and Asp25′ of said HIV aspartyl protease;

L¹ and L² are independently acyclic or cyclic linker moieties; and

each of Z¹ and Z² may be optionally present and, if present, areindependently selected from groups which occupy a volume of spaceoverlapping with the volume of space that would be filled by the nativesubstrate of said HIV aspartyl protease.

More preferred compounds of formula XL contain at least one group Q¹ orQ² comprising —SO₂—. Most preferrred compounds of formula XL contain atleast one group Q¹ or Q² comprising a substituted sulfonamide.

In one embodiment of this invention, compounds of formula XL may befurther constrained by “conformational locks”, such as a macrocyclicring structure. Such constraints are well known in the art ofpeptidomimetics and may result in compounds with strong biologicalactivity. See, for example, Dhanoa, D. S. et al. “The Synthesis ofPotent Macrocyclic Renin Inhibitors” Tetrahedron Lett. 33, 1725 (1992)and Flynn, G. A. et al. “An Acyl-Iminium Ion Cyclization Route to aNovel Conformationally Restricted Dipeptide Mimic: Applications toAngiotensin-Converting Enzyme Inhibition” J. Am. Chem. Soc. 109, 7914(1989)).

This invention also includes novel methods for accurate identification,design, or prediction of HIV inhibitors characterized by structural andphysicochemical features (1) through (6). By virtue of these methods,the skilled artisan can routinely predict and produce particularlyeffective HIV protease inhibitors.

We have found that the following method for identification, design orprediction of effective HIV protease inhibitors is particularly useful;

-   -   (a) selecting a candidate compound of defined chemical structure        containing a first and a second hydrogen bond acceptor moiety,        at least one of which is more highly polarizable than a        carbonyl, said moieties being the same or different; a third        hydrogen bonding moiety, which may be either a hydrogen bond        donor or acceptor; and at least two substantially hydrophobic        moieties;    -   (b) determining a low-energy conformation for binding of said        compound to the active site of an HIV aspartyl protease;    -   (c) evaluating the capability of said first and second hydrogen        bond acceptor moieties to form hydrogen bonds to the flap water        molecule of said HIV aspartyl protease when said compound is        bound thereto in said conformation;    -   (d) evaluating the capability of said substantially hydrophobic        moieties to associate with the P₁ and P₁′ binding pockets of        said HIV aspartyl protease when said compound is bound thereto        in said conformation;    -   (e) evaluating the capability of said third hydrogen bonding        moiety to form hydrogen bonds to Asp25 and Asp25′ of said HIV        aspartyl protease when said compound is bound thereto in said        conformation;    -   (f) evaluating the overlap of the occupied volume of said        compound when said compound is bound to said HIV aspartyl        protease in said conformation and the occupied volume of a        native substrate of HIV aspartyl protease or a nonhydrolyzable        isostere thereof, when said polypeptide is bound to said HIV        aspartyl protease;    -   (g) evaluating the deformation energy of binding of said        compound to said HIV aspartyl protease;    -   (h) evaluating the enthalpic contribution of the sum of all        electrostatic interactions between said compound and said HIV        aspartyl protease when said compound is bound thereto in said        conformation; and    -   (i) accepting or rejecting said candidate compound as an HIV        protease inhbitor based upon the determinations and evaluations        carried out in steps (b) through (h).

Using the novel combination of steps set forth in this screening method,the skilled artisan can advantageously avoid time consuming andexpensive experimentation to determine enzymatic inhibition activity ofparticular compounds. The method is also useful for facilitatingrational design of HIV protease inhibitors and anti-HIV viral agents,including therapeutic and prophylactic agents against HIV infection.Accordingly, the present invention relates to such inhibitors andanti-viral agents produced by the screening method described above.

A variety of conventional techniques may be used to carry out each ofthe above evaluations. Generally, these techniques involve determiningthe location and binding proximity of a given moiety, the occupiedvolume of space of a bound compound, the deformation energy of bindingof a given compound and electrostatic interaction energies. Examples ofconventional techniques useful in the above evaluations include: quantummechanics, molecular mechanics, molecular dynamics, Monte Carlosampling, systematic searches and distance geometry methods (G. R.Marshall, Ann. Ref. Pharmacol. Toxicol., 27, p. 193 (1987)). Specificcomputer software has been developed for use in carrying out thesemethods. Examples of programs designed for such uses include: Gaussian92, revision C (M. J. Frisch, Gaussian, Inc., Pittsburgh, Pa. ©1992);AMBER, version 3.0 (U. C. Singh, University of California at SanFrancisco, ©1992); QUANTA/CHARMM (Molecular Simulations, Inc.,Burlington, Mass. ©1992); and Insight II/Discover (Biosysm TechnologiesInc., San Diego, Calif. ©1992). These programs may be implemented, forinstance, using a Silicon Graphics workstation, IRIS 4D/35 or IBMRISC/6000 workstation model 550. Other hardware systems and softwarepackages will be known and of evident applicability to those skilled inthe art.

Additional analysis of the actual detailed interactions of the HIVprotease-inhibitor complex can be employed to ascertain morespecifically the binding associations between the enzyme and the boundinhibitor. Such analysis may be carried out, for example, by studying asolution of the complex by single- and multi-dimensional NMR techniques.Advantageously, the enzyme and/or the inhibitor may be enriched withstable isotopes such as ¹³C, ¹⁵N and ²H to more easily determine bindingconformation and proximity. Techniques, such as isotope editing, may beused to enhance the resolution with which the interactions are observed.

Either as an alternative or a supplemental analysis, the HIVprotease-inhibitor complex may be studied by single crystal X-raydiffraction. The process of determining the structures ofprotein/inhibitor complexes using the X-ray techniques described aboveis well known and has been used for many different complexes (see T. L.Blundel and L. N. Johnson, Protein Crystallography, Academic Press,(1976) and Methods in Enzymology, volumes 114 and 115, H. W. Wyckoff etal., eds., Academic Press (1985)). This technique can employ, forinstance, a highly purified preparation of HIV protease complexed withan inhibitor of interest in a buffered solution (typically at a pH ofbetween about 4.5 and about 8.0). The complex is allowed to crystallizein the presence of a precipitation agent (such as ammonium sulfate)under conditions which yield single crystals of the complex. Specificconditions for crystallizing HIV protease with various inhibitors havebeen well documented (see, for example, G. B. Dreyer et al.,Biochemistry, 31, p. 6646 (1992)). Application of a concentrated X-raybeam to an appropriately prepared and mounted crystal (preferably, anX-ray beam from a rotating anode X-ray generator or synchrotron) willyield a diffraction pattern from the reflected X-ray beam.

Detection of the diffracted rays may be carried out by visualizingphotographic paper exposed to the diffracted X-rays or alternatively, byusing a multiwire area detector (such as that manufactured by SiemensAnalytical X-Ray Instruments, Inc. (Madison, Wis.)) or an R-axis IIimage plate system from Rigaku Corporation (distributed by MolecularStructure Corporation, The Woodlands, Tex.). Other systems forgenerating and collecting X-ray diffraction data will be known to thoseof ordinary skill in the art.

Refinement of the X-ray diffraction data yields a three dimensionalstructure. Computer software (such as X-PLOR (Yale University, ©1992,distributed by Molecular Simulations, Inc.) has been developed to carryout this refinement.

In general, using the above techniques with an appropriately preparedcrystalline complex, a structure may be refined to about 2-3 Å with an Rvalue of about 0.25 or less. As the skilled artisan can appreciate,these values are adequate to determine the interactions between HIVprotease and a given compound such that it will be clear if features (1)through (6) are present and consequently, whether that given compound isan HIV aspartyl protease inhibitor. Thus, additional inhibitorsaccording to this invention may be designed and predicted based on acombination of crystallographic structural information and computationalanalysis.

For example, to predict the binding of a candidate inhibitor accordingto this invention, the inhibitor is examined to determine whether themolecule contains functionality which is not well represented by theexisting forcefield models in CHARMM (Molecular SimulationsIncorporated, Burlington, Mass.) or AMBER (Professor P. A. Kollman,UCSF). If any functionality is not well represented, we then examine allpublished structural information for molecules containing suchfunctionality, and in some cases perform high-level ab initiocalculations on simple molecules containing these functionalities todetermine their preferred conformations and the energy differencesbetween various conformations. More accurate parameters describing thesefunctional groups may then be derived for the CHARMM and/or AMBERforcefields and used in subsequent calculations.

Next, the candidate inhibitor is aligned in 3-dimensional space withother, related inhibitors whose bound conformations have previously beendetermined by x-ray crystallography. Both Van der Walls volume andelectrostatic potentials are used to direct the alignment process. Thealignment is typically done with software like Quanta (MolecularSimulations) or InsightII (Biosym Technologies, San Diego, Calif.). Thisalignment can be done manually within this software, or more automatedalignment procedures within the software (e.g. the “superimpose” optionof Quanta or the “APEX” module of InsightII) may be used. The result ofthis alignment is a first guess of the “bound” conformation of thecandidate inhibitor. This inhibitor is then docked in the active site ofHIV protease, and the confomation is energy minimized with the enzymeatoms held fixed in space. These minimizations are typically done usingthe CHARMM or AMBER forcefields.

Because inhibitors can sometimes bind in multiple or unexpectedconformations within an active site, we often then carry out furthersearches of the bound conformation of the enzyme-inhibitor complex. Forexample, a variety of Monte Carlo search techniques (e.g. as found inthe Conformational Search Module of Quanta) may be used, along withhigh-temperature dynamics and simulated annealing. These searchtechniques reveal whether there are alternative, reasonable low-energyconfomrations in which the inhibitor may bind to the enzyme. The effectsof solvation and desolation in the formation of the variousenzyme-inhibitor complexes may be estimated with programs such as DELPHI(Biosym), Polaris (Molecular Simulations) and AMSOL (Professor C.Cramer, University of Minnesota). The result of this searching is a setof one or more bound conformations for the candidate inhibitor.

For each of the low-energy conformations, waters may then be added tothe active site of the enzyme and the entire system relaxed. Finally,molecular dynamics simulations may be used to study the detailed motionsof the enzyme, the inhibitor, and related water molecules.

The final set of remaining low-energy conformations (typically a verysmall number) represents our predictions of the bound conformation ofthe candidate inhibitor. Each conformation includes our estimate of thedynamic flexibility of the entire system (inhibitor, enzyme, andwaters).

The more advanced methodology is typically applied to the study of thefirst few compounds in a series, when there are the greatestuncertainties about the possible binding mode(s) in the enzyme activesite. For later compounds within a series, the low energy conformersobtained from the searches on earlier compounds provide informationabout the possible low energy conformers of the inhibitor compounds. Inaddition, crystallographic information about the conformation of thebound complexes of earlier compounds within a series is often available.This prior computational and structural work advantageously facilitatesthe prediction of the bound conformation of candidate inhibitormolecules.

To exemplify the above screening method, we have carried out thefollowing evaluation of compound 140 (Table II), a preferred compound ofthis invention, as described below.

Prediction of Binding Conformation and Energy of Compound 140 to HIVProtease

The forcefield for the benzenesulfonamide portion of compound 140 wasderived from ab initio calculations and incorporated into the AMBERforcefield. The latest CHARMM forcefield parameters for this moiety werefound to be adequate for energy minimization studies and are used in allQuanta/CHARMM calculations.

The low energy conformers obtained from the conformational searches onearlier compounds in the sulfonamide series (such as compound 16)provided information about the possible low energy conformers ofcompound 140. These low energy conformers were aligned in 3-dimensionalspace with other related inhibitors whose bound conformations havepreviously been determined by x-ray crystallography. This alignmentprocess was carried out manually within Quanta and, in some cases, wasassisted with the “conformational search” option of Quanta. Thereference crystal structure used in this alignment was the complex ofHIV-1 protease with compound 16. This inhibitor structure was energyminimized in the active site of the enzyme using Quanta/CHARM. Theenzyme atoms were held fixed during this minimization. Only the flapwater was included. Later simulations allowed the enzyme to relax andused a variety of dielectric approximations. A single low-energyconformation which was consistent with all previous conformationalsimulations and crystallographic data was obtained (see FIG. 1). Thispredicted binding conformation was later found to be essentially inagreement with the results obtained by x-ray crystallography (see FIGS.2 and 3).

As discussed above, the novel compounds of the present invention areexcellent ligands for aspartyl proteases, particularly HIV-1 and HIV-2proteases. Accordingly, these compounds are capable of targeting andinhibiting late stage events in HIV replication, i.e., the processing ofthe viral polyproteins by HIV encoded proteases. Such compounds inhibitthe proteolytic processing of viral polyprotein precursors by inhibitingaspartyl protease. Because aspartyl protease is essential for theproduction of mature virions, inhibition of that processing effectivelyblocks the spread of virus by inhibiting the production of infectiousvirions, particularly from chronically infected cells. Compoundsaccording to this invention advantageously inhibit the ability of theHIV-1 virus to infect immortalized human T cells over a period of days,as determined by an assay of extracellular p24 antigen—a specific markerof viral replication. Other anti-viral assays have confirmed the potencyof these compounds.

The compounds of this invention may be employed in a conventional mannerfor the treatment of viruses, such as HIV and HTLV, which depend onaspartyl proteases for obligatory events in their life cycle. Suchmethods of treatment, their dosage levels and requirements may beselected by those of ordinary skill in the art from available methodsand techniques. For example, a compound of this invention may becombined with a pharmaceutically acceptable adjuvant for administrationto a virally-infected patient in a pharmaceutically acceptable mannerand in an amount effective to lessen the severity of the viralinfection.

Alternatively, the compounds of this invention may be used in vaccinesand methods for protecting individuals against viral infection over anextended period of time. The compounds may be employed in such vaccineseither alone or together with other compounds of this invention in amanner consistent with the conventional utilization of proteaseinhibitors in vaccines. For example, a compound of this invention may becombined with pharmaceutically acceptable adjuvants conventionallyemployed in vaccines and administered in prophylactically effectiveamounts to protect individuals over an extended period time against HIVinfection. As such, the novel protease inhibitors of this invention canbe administered as agents for treating or preventing HIV infection in amammal.

The compounds of formula I, especially those having a molecular weightof less than about 700 g/mole, may be readily absorbed by thebloodstream of mammals upon oral administration. Compounds of formula Ihaving a molecular weight of less than about 600 g/mole are most likelyto demonstrate oral availability. This surprisingly impressive oralavailability makes such compounds excellent agents fororally-administered treatment and prevention regimens against HIVinfection.

The compounds of this invention may be administered to a healthy orHIV-infected patient either as a single agent or in combination withother anti-viral agents which interfere with the replication cycle ofHIV. By administering the compounds of this invention with otheranti-viral agents which target different events in the viral life cycle,the therapeutic effect of these compounds is potentiated. For instance,the co-administered anti-viral agent can be one which targets earlyevents in the life cycle of the virus, such as cell entry, reversetranscription and viral DNA integration into cellular DNA. Anti-HIVagents targeting such early life cycle events include, didanosine (ddI),alcitabine (ddC), d4T, zidovudine (AZT), polysulfated polysaccharides,sT4 (soluble CD4), ganiclovir, dideoxycytidine, trisodiumphosphonoformate, eflornithine, ribavirin, acyclovir, alpha interferonand trimenotrexate. Additionally, non-nucleoside inhibitors of reversetranscriptase, such as TIBO or nevirapine, may be used to potentiate theeffect of the compounds of this invention, as may viral uncoatinginhibitors, inhibitors of trans-activating proteins such as tat or rev,or inhibitors of the viral integrase.

Combination therapies according to this invention exert a synergisticeffect in inhibiting HIV replication because each component agent of thecombination acts on a different site of HIV replication. The use of suchcombinations also advantageously reduces the dosage of a givenconventional anti-retroviral agent which would be required for a desiredtherapeutic or prophylactic effect as compared to when that agent isadministered as a monotherapy. These combinations may reduce oreliminate the side effects of conventional single anti-retroviral agenttherapies while not interfering with the anti-retroviral activity ofthose agents. These combinations reduce potential of resistance tosingle agent therapies, while minimizing any associated toxicity. Thesecombinations may also increase the efficacy of the conventional agentwithout increasing the associated toxicity. In particular, we havediscovered that these compounds act synergistically in preventing thereplication of HIV in human T cells. Preferred combination therapiesinclude the administration of a compound of this invention with AZT,ddI, ddC or d4T.

Alternatively, the compounds of this invention may also beco-administered with other HIV protease inhibitors such as Ro 31-8959(Roche), L-735,524 (Merck), XM 323 (Du-Pont Merck) and A-80,987 (Abbott)to increase the effect of therapy or prophylaxis against various viralmutants or members of other HIV quasi species.

We prefer administering the compounds of this invention as single agentsor in combination with retroviral reverse transcriptase inhibitors, suchas derivatives of AZT, or other HIV aspartyl protease inhibitors. Webelieve that the co-administration of the compounds of this inventionwith retroviral reverse transcriptase inhibitors or HIV aspartylprotease inhibitors may exert a substantial synergistic effect, therebypreventing, substantially reducing, or completely eliminating viralinfectivity and its associated symptoms.

The compounds of this invention can also be administered in combinationwith immunomodulators (e.g., bropirimine, anti-human alpha interferonantibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha,diethyldithiocarbamate, tumor necrosis factor, naltrexone and rEPO); andantibiotics (e.g., pentamidine isethiorate) to prevent or combatinfection and disease associated with HIV infections, such as AIDS andARC.

When the compounds of this invention are administered in combinationtherapies with other agents, they may be administered sequentially orconcurrently to the patient. Alternatively, pharmaceutical orprophylactic compositions according to this invention may be comprisedof a combination of an aspartyl protease inhibitor of this invention andanother therapeutic or prophylactic agent.

Although this invention focuses on the use of the compounds disclosedherein for preventing and treating HIV infection, the compounds of thisinvention can also be used as inhibitory agents for other viruses whichdepend on similar aspartyl proteases for obligatory events in their lifecycle. These viruses include, as well as other AIDS-like diseases causedby retroviruses, such as simian immunodeficiency viruses, but are notlimited to, HTLV-I and HTLV-II. In addition, the compounds of thisinvention may also be used to inhibit other aspartyl proteases, and inparticular, other human aspartyl proteases, including renin and aspartylproteases that process endothelin precursors.

Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle. Pharmaceutically acceptable carriers, adjuvants and vehiclesthat may be used in the pharmaceutical compositions of this inventioninclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. We prefer oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. The termparenteral as used herein includes subcutaneous, intracutaneous,intravenous, intramuscular, intra-articular, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant such as Ph. Helv or a similar alcohol.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carriers which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried corn starch. Whenaqueous suspensions are administered orally, the active ingredient iscombined with emulsifying and suspending agents. If desired, certainsweetening and/or flavoring and/or coloring agents may be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible by topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxy-ethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esterswax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable enema formulation. Topically-transdermal patches arealso included in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art.

Dosage levels of between about 0.01 and about 100 mg/kg body weight perday, preferably between about 0.5 and about 50 mg/kg body weight per dayof the active ingredient compound are useful in the prevention andtreatment of viral infection, including HIV infection. Typically, thepharmaceutical compositions of this invention will be administered fromabout 1 to about 5 times per day or alternatively, as a continuousinfusion. Such administration can be used as a chronic or acute therapy.The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. A typicalpreparation will contain from about 5% to about 95% active compound(w/w). Preferably, such preparations contain from about 20% to about 80%active compound.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease.Patients may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

As the skilled artisan will appreciate, lower or higher doses than thoserecited above may be required. Specific dosage and treatment regimensfor any particular patient will depend upon a variety of factors,including the activity of the specific compound employed, the age, bodyweight, general health status, sex, diet, time of administration, rateof excretion, drug combination, the severity and course of theinfection, the patient's disposition to the infection and the judgmentof the treating physician.

The compounds of this invention are also useful as commercial reagentswhich effectively bind to aspartyl proteases, particularly HIV aspartylprotease. As commercial reagents, the compounds of this invention, andtheir derivatives, may be used to block proteolysis of a target peptideor may be derivatized to bind to a stable resin as a tethered substratefor affinity chromatography applications. These and other uses whichcharacterize commercial aspartyl protease inhibitors will be evident tothose of ordinary skill in the art.

In order that this invention be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

General Materials and Methods

All temperatures are recorded in degrees Celsius. Thin layerchromatography (TLC) was carried out using 0.25 mm thick E. Merck silicagel 60 F₂₅₄ plates and elution with the indicated solvent system.Detection of the compounds was carried out by treating the plate with anappropriate visualizing agent, such as 10% solution of phosphomolybdicacid in ethanol or a 0.1% solution of ninhydrin in ethanol, followed byheating, and/or by exposure to UV light or iodine vapors whenappropriate. Thick layer silica gel chromatography was also carried outusing E. Merck 60 F₂₅₄ plates “prep plates”) of 0.5, 1.0, or 2.0 mmthickness. Following development of the plate, the band of silicacontaining the desired compound was isolated and eluted with anappropriate solvent. Analytical HPLC was carried out using a Water'sDelta Pak, 5 μM silica, C18 reversed-phase column, 3.9 mm ID×15 cm Lwith a flow rate of 1.5 mL/min using the following table:

Mobile phase: A = 0.1% CF₃CO₂H in H₂O B = 0.1% CF₃CO₂H in CH₃CNGradient: T = 0 min., A (95%), B (5%) T = 20 min., A (0%), B (100%) T =22.5 min., A (0%), B (100%)Preparative HPLC was also carried out using C₁₈ reversed-phase media.HPLC retention times were recorded in minutes. NMR spectral data wasrecorded using a Bruker AMX500, equipped with either a reverse or QNPprobe, at 500 MHz, and was taken in the indicated solvent.

We have measured the inhibition constants of each compound against HIV-1protease using the method described essentially by M. W. Pennington etal., Peptides 1990, Gimet, E. and D. Andrew, Eds., Escom; Leiden,Netherlands (1990).

Compounds of formula I were tested for their antiviral potency inseveral virological assays. In the first assay, the compounds were addedas a solution in dimethylsulfoxide (DMSO) to a test cell culture ofCCRM-CEM cells, a strain of CD4⁺ human T-cell lymphoma cells, previouslyacutely infected with HIV_(IIIb) using standard protocols (see Meek, T.D. et al., “Inhibition of HIV-1 protease in infected T-lymphocytes bysynthetic peptide analogues”, Nature, 343, p. 90 (1990). Preferredcompounds are those which are able to inhibit 90% of viral infectivityat a concentration of 1 μM or less. More preferred compounds are thosewhich are able to inhibit 90% of viral infectivity at a concentration of100 nM or less.

The effect of the compounds on inhibiting the replication of the viruswas measured by determining the HIV extracellular p24 antigenconcentration using a commercial enzyme immunoassay (obtained fromCoulter Corporation, Hialeah, Fla.).

Depending on the cell type and the desired readout, syncytia formation,reverse-transcriptase (RT) activity, or cytopathic effect as assayed bya dye uptake method may also be used as readouts of antiviral activity.See H. Mitsuya and S. Broder, “Inhibition of the in vitro infectivityand cytopathic effect of human T-lymphotropic virus typeIII/lymphoadenopathy-associated virus (HTLV-III/LAV) by2′,3′-dideoxynucleosides”, Proc. Natl. Acad. Sci. USA, vol. 83, pp.1911-1915 (1986). The effect of compounds of formula I on clinicalisolates of other HIV-1 strains was determined by obtaining low-passagedvirus from HIV-infected patients and assaying the effect of theinhibitors in preventing infection of the HIV virus in freshly preparedhuman peripheral blood mononuclear cells (PBMCs).

Insofar as compounds of formula I are able to inhibit the replication ofthe HIV virus in human T-cells and furthermore, may be delivered orallyto mammals, they are of evident clinical utility for the treatment ofHIV infection. These tests are predictive of the compounds ability toinhibit HIV protease in vivo.

EXAMPLE 1

-   A. Compound XI ((syn) —OH, D′=benzyl). 184 g of Brockman Super I    grade neutral alumina was slurried in sufficient diethyl ether to    form a thick, stirrable suspension and was treated with 7.48 mL of    benzylamine. After stirring for 5 min, 7.28 g of    (1S,2S)-1-(N-benzyoxycarbonyl)-amino-2-phenylethyl-oxirane was added    and the mixture stirred for 15 h. The mixture was treated with 15.28    g of di-tert-butylpyrocarbonate and 4.70 mL of    diisopropylethylamine. This mixture was stirred for 3.5 h, then    treated with 600 mL of methanol, allowed to stand for 3.5 h, and    filtered to yield a yellow oil, which was purified by silica gel    chromatography using a gradient of 0.5 to 1.5% methanol in methylene    chloride to yield 3.88 g of the desired product as a white solid.    Further washing the filter cake with methanol and with 3% ammonium    hydroxide in methanol yielded 2.2 g of    4-benzylamino-2-N-benzyloxycarbonylamino-3-hydroxy-1-phenylbutane in    several portions. Each of these portions was treated separately, as    a solution in methylene chloride, with 1.1 molar equivalents each of    di-tert butylpyrocarbonate and diisopropylethylamine, followed by    aqueous workup with water, 10% aqueous KHSO₄, and brine, drying over    MgSO₄, and concentration in vacuo. The combined products of these    reactions were purified by silica gel chromatography using a    gradient of 5% to 15% diethyl ether in methylene chloride. The    resulting pure fractions were collected and combined with the    previously purified product to yield 5.49 g of a white solid. TLC:    Rf=0.56, 5% methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound XII ((syn) —OH, D′=benzyl). A solution of 5.49 g of the    resultant compound of Example 1A in 40 mL of ethanol was    hydrogenated under a slight positive pressure of hydrogen in the    presence of 380 mg of 10% palladium on carbon for 16 h. After    filtering and concentrating in vacuo, the desired product was    obtained as 4.03 g of a white solid. TLC: Rf=0.21, 95:5:0.5    CH₂Cl₂/methanol/concentrated NH₄OH.-   C. Compound XIII ((syn) —OH, A=benzyloxycarbonyl, D′=benzyl). A    solution of 3.02 g of the resultant compound of Example 1B in 150 mL    of methylene chloride was treated with 4.35 g of    N^(α)-Cbz-N^(δ)-trityl asparagine, 1.16 g of hydroxybenzotriazole    hydrate, and 1.64 g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide    hydrochloride. The mixture was stirred for 16 h, then diluted with 3    volumes of diethyl ether and washed sequentially with water,    saturated NaHCO₃ solution, 10% KHSO₄ solution, and brine. After    drying over MgSO₄ and concentrating in vacuo, a yellow oil was    obtained which was purified by chromatography on a Florisil column    using a gradient of 0% to 25% EtOAc in CH₂Cl₂ as eluant to yield    8.00 g of the title compound as a white foam. TLC: Rf=0.51, 5%    methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.-   D. Compound XIV ((syn) —OH, A=H, D′=benzyl). A solution of 7.90 g of    the resultant compound of Example 1C in 150 mL of ethanol was    hydrogenated under a slight positive pressure of hydrogen int he    presence of 550 mg of 10% palladium on carbon for 2.5 h, then ca. 50    mg more 10% palladium on carbon was added, the mixture was then    filtered and concentrated in vacuo to give the desired product as    6.66 g of a white solid which was used without subsequent    purification. TLC: Rf=0.26, 95:5:0.5 CH₂Cl₂/methanol/concentrated    NH₄OH.-   E. Compound XIV ((syn) —OH, A=quinoline-2-carbonyl, D′=benzyl). A    suspension of 1.51 g of quinaldic acid and 6.17 g of the resultant    compound of Example 1D in 150 mL of acetonitrile was treated with    1.52 mL of diisopropylethylamine and 3.58 g of BOP reagent. The    mixture was stirred for 14 h, then concentrated in vacuo. The gummy    residue was partitioned between ether and water, and the organic    layer was washed sequentially with brine, saturated NaHCO₃ solution,    water, 10% KHSO₄ solution, and brine, then dried over MgSO₄ and    concentrated in vacuo. Subsequent purification by silica gel    chromatography using 0% to 8.5% solvent A in methylene chloride    (where solvent A is defined as 90:10:1, methylene    chloride/methanol/concentrated ammonium hydroxide) yielded 5.79 g of    the title compound as a white foam, along with ca. 600 mg of    slightly impure side fractions. TLC: Rf=0.41, 5% methanol/CH₂Cl₂;    (¹H)-NMR (CDCl₃) consistent with structure.-   F. Compound 1. A 58 mg portion of the resultant compound of Example    1E was treated with 1 mL of 90% aqueous TFA and allowed to stand for    17 h. The mixture was concentrated in vacuo and the residue taken up    in 3 mL of CH₂Cl₂, treated with 100 μL of DIEA, and cooled to 0° C.    To this solution was added 26 μL of benzenesulfonyl chloride, and    the mixture was stirred for 18 h, warming slowly to ambient    temperature. After concentration of the mixture in vacuo, the    residue was purified by thick layer silica gel chromatography using    5% MeOH/CH₂Cl₂ as eluant followed by preparative reversed-phase C₁₈    HPLC using a linear gradient of 40% to 100% CH₃CN/H₂O with 0.1% TFA    for elution to obtain 8.3 mg of the title compound. TLC: Rf=0.50, 5%    MeOH/CH₂Cl₂. HPLC: Rt=17.8 min. NMR (DMSO-d₆) δ 2.62 (dd, 1H); 2.76    (d, 2H); 2.80 (dd, 1H); 3.11, (d, 2H); 3.34 (dd, 1H); 4.59 (br s,    1H); 4.68 (br s, 1H); 3.97 (m, 1H); 4.20 (d, 1H), 4.35 (d, 1H); 4.68    (dd, 1H); 6.39 (d, 1H); 6.74 (t, 1H); 6.81 (t, 2H); 6.93 (d, 2H);    7.12-7.24 (m, 6H); 7.51 (t, 2H); 7.57 (t, 1H); 7.62 (dd, 1H); 7.77    (t, 2H), 7.96 (d, 1H); 8.09 (d, 1H); 8.16 (d, 1H); 8.31 (d, 1H);    8.53 (d, 1H).

EXAMPLE 2

Compound 2. A 150 mg portion of the resultant compound of Example 1E wasdissolved in 1 mL of 90% aqueous TFA and stirred at ambient temperatureovernight, then concentrated in vacuo. The crude TFA salt residue wasdissolved in 7 mL of dry methylene chloride and the pH of the solutionwas adjusted to pH 8 with 1N NaOH. 56 mg of a mixture of4-fluoro-3-acetamidobenzene sulfonylchloride and3-fluoro-4-acetamidobenzene sulfonylchloride (˜1:1) was added and themixture stirred vigorously for 3 hours after which an additional 25 mgwas added and the reaction allowed to continue for an additional 12hours. The reaction was then diluted with 50 mL of ethylene chloride andthe organic layer was washed sequentially with water and brine, driedover MgSO₄ and concentrated in vacuo. The crude residue was purifiedusing a silica gel flash chromatography column using a gradient of 3% to5% MeOH in methylene chloride as eluant to yield 60 mg of the titlecompounds. TLC: Rf=0.50, 10% MeOH/CH₂Cl₂; HPLC: Rt=13.93 min. NMR(CDCl₃): δ9.05 (s, 1H); 8.65 (d, 0.5H); 8.58 (t, 0.5H), 8.20 (dd, 0.5H),7.85 (d, 1H) 7.75 (m, 0.5H), 7.45-7.63 (m, 1.5H), 7.14-7.25 (m, 6H),6.78-6.95 (m, 5H), 6.70 (d, 1H), 6.41 (s, 0.5H), 6.25 (s, 0.5H), 6.18(s, 0.5H), 6.10 (s, 0.5H), 4.88 (m, 0.5H), 4.81 (m, 0.5H), 4.37 (d, 1H),4.35 (m, 1H), 4.21 (d, 1H), 4.00 (m, 1H), 3.46 (m, 0.5H), 3.35 (m,0.5H), 3.27 (d, 0.5H), 3.16 (d, 0.5H), 3.14 (d, 1H), 2.45-2.75 (m, 5H);2.16, 2.20 (2 s, 3H total).

EXAMPLE 3

Compound 3. A 23 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 15 h. Themixture was concentrated in vacuo and the residue taken upon in 2 mL ofCH₂Cl₂, treated with 6 μL of DIEA, and cooled to 0° C. To this solutionwas added 23 mg of 3,5-dimethylisoxazole-4-sulfonyl chloride, and themixture was stirred for 18 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 1.1 mg of the titlecompound. TLC: Rf=0.55, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.5 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 4

Compound 4. A 33 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 15 h. Themixture was concentrated in vacuo and the residue taken up in 3 mL ofCH₂Cl₂, treated with 16 μL of DIEA, and cooled to 0° C. To this solutionwas added 10 μL of 3-trifluoromethylbenzene sulfonyl chloride, and themixture was stirred for 18 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 1.1 mg of the titlecompound. TLC: Rf=0.55, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.5 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 5

Compound 5. A 20 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 18 h. Themixture was concentrated in vacuo and the residue taken up in 1 mL ofCH₂Cl₂, treated with 10 μL of DIEA, and cooled to 0° C. To this solutionwas added 13 mg of 2-acetamido-4-methyl-5-thiazolesulfonyl chloride, andthe mixture was stirred for 17 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 0.40 mg of the titlecompound. TLC: Rf=0.5, 10% MeOH/CH₂Cl₂. HPLC: Rt=13.8 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 6

Compound 6. A 33 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 16 h. Themixture was concentrated in vacuo and the residue taken up in 2 mL ofCH₂Cl₂, treated with 16 μL of DIEA, and cooled to 0° C. To this solutionwas added 11 mg of 5-(isoxazol-3-yl)thiophene-2-sulfonyl chloride, andthe mixture was stirred for 18 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 1.5 mg of the titlecompound. TLC: Rf=0.7, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.7 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 7

Compound 7. A 35.5 mg portion of the resultant compound of Example 1Ewas treated with 1 mL of 90% aqueous TFA and allowed to stand for 18 h.The mixture was concentrated in vacuo and the residue taken up in 3 mLof CH₂Cl₂, treated with 16 μL of DIEA, and cooled to 0° C. To thissolution was added 10 mg of 3-chlorosulfonylbenzoic acid, and themixture was stirred for 16 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% tTFA for elution to obtain 1.6 mg of the titlecompound. TLC: Rf=0.7, 10% MeOH/CH₂Cl₂. HPLC: Rt=13.6 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 8

Compound 8. 0.04 mmol of the resultant compound of Example 10A wasconverted to the free base by partitioning between EtOAc and sat.NaHCO₃. Treatment of the resulting compound with an excess of 1%HCl/MeOH and concentration in vacuo yielded the hydrochloride salt as awhite solid. This compound was suspended in CH₂Cl₂ and treated withsufficient DIEA to bring the pH to >10 (moist pH paper). The solutionwas treated with 7 molar equivalents of chlorotrimethylsilane andstirred for 15 h under nitrogen, then treated with 0.06 mmol of methanesulfonyl chloride and stirred for 1 h. The resulting mixture wasconcentrated to a small volume, applied directly to a thick layer silicagel plate and eluted with 7% MeOH/CH₂Cl₂. The primary UV-quenching bandwas isolated and further purified by preparative reversed-phase HPLC toyield the title compound as a white solid. TLC: Rf=0.65, 10%CH₃OH/CH₂Cl₂, HPLC: Rt=12.3 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLES 9 and 192

-   A. Compound XIV ((syn, anti-OH, A=quinoline-2-carbonyl,    D′=isobutyl). A solution of 317 mg (0.425 mmol) of the resultant    compounds of Example 17B, diastereomer B and 0.11 mL (0.637 mmol) of    diisopropylethyl amine in 7 mL of dichloromethane was treated with    139.1 mg (0.637 mmol) of di-tert-butyl dicarbonate. After 24 hours,    the mixture was diluted with dichloromethane. The mixture was washed    with water, 5% NaHCO₃, 0.5 N HCl, brine then dried over MgSO₄,    filtered and concentrated in vacuo. The residue was purified by low    pressure silica gel column chromatography using a 20% ethyl    acetate/dichloromethane as eluent to yield 81.2 mg of the fast    moving hydroxyl diastereomer, 65.8 mg of the slower moving hydroxyl    diastereomer, and 65.8 mg of the mixed diastereomers. TLC: Rf=0.60,    0.67, 40% EtOAc/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compounds 9 and 192. A solution of 35.1 mg (0.041 mmol) of the    resultant mixed diastereomers (˜1:1) of Example 9/192A in 0.8 mL of    dichloromethane was treated with 0.8 mL of trifluoroacetic acid.    After 4 hours, the mixture was concentrated in vacuo. TLC: Rf=0.11,    10% CH₃OH/CH₂Cl₂. To a solution of the resulting trifluoroacetic    acid salt (entire yield) in 1 mL of dichloromethane was    sequentialled added 0.3 mL of saturated NaHCO₃, a small amount of    solid NaHCO₃ and 11.8 mg (0.054 mmol) of benzofurazan-4-sulphonyl    chloride. After 3 hours, the mixture was diluted with    dichloromethane. The two layers were separated and the aqueous layer    was extracted once with dichloromethane. The combined organic layer    was washed with brine then dried over MgSO₄, filtered and    concentrated in vacuo. The residue was purified by preparative HPLC    to yield 2.0 mg of compound 9 as a white solid: TLC: Rf=0.20, 5%    CH₃OH/CH₂Cl₂; HPLC, RT 14.2 min. 2.7 mg of compound 192 was also    obtained as a white solid, which was determined by NMR and HPLC to    be contaminated with ˜25% of compound 9: TLC: Rf=0.20, 5%    CH₃OH/CH₂Cl₂; HPLC, Rt=14.2 min. (¹H)-NMR consistent with structure.

EXAMPLE 10

-   A. Compound XV ((syn) —OH, A=quinoline-2-carbonyl, D′=benzyl; TFA    salt). A 0° C. solution of 1.027 g portion of the resultant compound    of Example 1E in 5 mL of CH₂Cl₂ was treated with 5 mL of TFA and    allowed to stand for 3 h. The mixture was concentrated in vacuo to    yield 0.95 g of the title compound, which was used without    subsequent purification.-   B. Compound 10. A solution of 30.2 mg of the resultant compound of    Example 10A in 3 mL of CH₂Cl₂ was treated with 0.33 mL of DIEA and    31.1 mg of m-benzenedisulfonyl chloride. The mixture was stirred for    2 h, then treated with 2 mL of concentrated aqueous ammonium    hydroxide. The biphasic mixture was stirred for an additional 16 h,    concentrated in vacuo, and the residue partitioned between ethyl    acetate and brine. The organic layer was dried over anhydrous MgSO₄    and concentrated in vacuo, and the residue was purified by    preparative thick layer silica gel chromatography using 3%    MeOH/CH₂Cl₂ as eluant to yield 4.5 mg of the title compound. TLC:    Rf=0.5, 3% MeOH/CH₂Cl₂ as eluant to yield 4.5 mg of the title    compound. TLC: Rf=0.5, 3% MeOH/CH₂Cl₂. HPLC: Rt=13.4 min; (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 11

Compound 11. A solution of 57.9 mg of the resultant compound of Example10A in 5 mL of CH₂Cl₂ was treated with 30 μL of DIEA and 9.3 μL ofdimethylsulfamoyl chloride. The mixture was stirred for 12 h, thentreated with an additional 30 μL of DIEA and 9.3 μL of dimethylsulfamoylchloride and the reaction was allowed to proceed an additional 12 hours.The mixture was then diluted with CH₂Cl₂ and washed with saturatedNH₄Cl; the aqueous layer was washed with CH₂Cl₂, and the combinedorganic extracts were dried over MgSO₄. Filtration and concentrationprovided a residue which was chromatographed on a silica gel columnusing 2.5% MeOH/EtOAc as eluent, yielding a slightly impure productwhich was further purified by preparative HPLC using a linear gradientof 35% to 100% CH₃/CN/H₂O with 0.1% TFA for elution. HPLC: Rt=13.0minutes. NMR (CDCL₃): δ9.15 (d, 1H), 8.34 (d, 1H), 8.22 (d, 1H), 8.18(d, 1H), 7.90 (d, 1H), 7.80 (t, 1H), 7.65 (t, 1H), 7.16-738 (m, 5H),7.05 (d, 1H), 6.95 (t, 1H), 6.87 (t, 1H), 5.85 (br s, 1H), 5.62 (br s,1H), 4.87 (M, 1H), 4.46 (s, 2H), 4.08 (m, 1H), 3.66 (m, 1H), 3.30 (m,2H), 2.59-2.94 (m, 4H), 2.81 (s, 6H).

EXAMPLE 12

-   A. Compound XIV ((syn) —OH, A=quinoline-2-carbonyl, D′=benzyl;    trifluoroacetate salt). To a solution of 1.027 g (1.164 mmol) of the    resultant compound of Example 1E in CH₂Cl₂ (5 mL) at 0° to 5° C. was    added trifluoromethanesulfonic acid (5 mL). After stirring for 3 h,    the reaction mixture was concentrated in vacuo to provide 0.95 g of    light yellow, gummy product, containing one equivalent. of    triphenylmethanol, which was used without subsequent purification.-   B. Compound 12. To a solution of 30.2 mg (0.038 mmol) of the    resultant compound of Example 12A in CH₂Cl₂ (3 mL) was added    diisopropylethylamine (0.33 mL, 0.189 mmol), and    2-(pyrid-2-yl)-tyiophene-5-sulfonyl chloride 13 mg, (0.249 mmol).    After 14 h, the resulting mixture was diluted with ethyl acetate,    washed with saturated brine, dried over magnesium sulfate, filtered    and concentrated in vacuo. The residue was purified by preparative    reversed-phase chromatography using a 5% to 100% H₂O/acetonitrile    gradient as eluant to yield the title product.

EXAMPLE 13

Compound 13. To a solution of 30 mg (0.038 mmol) of the resultantcompound of Example 12A in CH₂Cl₂ (3 mL) was added diisopropylethylamine(0.33 mL, 0.189 mmol), and 2-(3-phenylsulfonyl)thiophene sulfonylchloride (0.113 mmol). After stirring for 2 h, the reaction mixture wasmade biphasic by addition of 30% ammonium hydroxide solution (2 mL).After stirring for an additional 16 h, the resultant mixture wasconcentrated in vacuo, reconstituted in ethyl acetate, washed withsaturated brine, dried over magnesium sulfate, filtered, andre-concentrated in vacuo. Purification by thin layer preparativechromatography yielded the desired compound.

EXAMPLE 14

Compound 14. The resulting compound of Example 17B, diastereomer B (170mg) was treated with 1 mL of 90% aqueous TFA and allowed to stand for 12h. The mixture was concentrated in vacuo and the residue taken up in 5mL of dry CH₂Cl₂. To this solution, 3 mL of saturated aqueous sodiumbicarbonate and 50 mg of 4-fluorobenzenesulfonyl chloride was added andthe mixture stirred for 3 h. The resulting mixture was diluted withCH₂Cl₂ and washed with water, dried over magnesium sulfate and filtered.After concentration of the mixture in vacuo, a portion of the residuewas purified by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA for elution to obtain3.0 mg of the title compound. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC:Rt=14.78 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 15

Compound 15. A sample of a mixture of4-fluoro-3-acetamidobenzenesulfonyl chloride and3-fluoro-4-acetamidobenzenesulfonyl chloride (approx. 1:1; obtained fromMaybridge Chemicals) was resolved into its respective regioisomers bysilica gel chromatography using 10% isopropyl alcohol/hexane as eluent.A solution of 4-acetamido-3-fluorobenzenesulfonyl chloride (30 mg) andthe resulting compound of Example 17B, diastereomer B (80 mg) in 10 mLof CH₂Cl₂ was reacted in the same manner as described for Example 14.After workup and purification of a portion of the product by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1 TFA as eluent, 1.2 mg of the title compound was obtained as awhite solid. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC: Rt=12.91 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 16

Compound 16. 80 mg of the resulting compound of Example 17B,diastereomer B, was reacted with 45 mg of3-acetamido-4-fluorobenzenesulfonyl chloride in the same manner asdescribed for Example 14. After workup and purification of a portion ofthe product by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 1.4 mg of thetitle compound was obtained. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC:Rt=12.91 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 17

-   A. (2S)-2-((1S, 2R syn,    anti)-3-(2-methylpropyl)amino-1-benzyl-2-hydoxypropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-trityl    succinamide. A solution of 683.1 mg (0.96 mmol) of the resultant    compounds of Example 191D and 1.9 mL (19.2 mmol) of isobutylamine in    10 mL of acetonitrile in a sealed tube was heated at 90-100° C. for    24 hours. After cooling to room temperature, the mixture was    concentrated in vacuo. The residue was taken up in dichloromethane    and washed with water, brine, then dried over MgSO₄, filtered and    concentrated in vacuo to yield 783.8 mg of the mixed diastereomeric    products. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XIII, ((syn, anti) —OH, P=quinoline-2-carbonyl,    D′=isobutyl). A solution of 583.8 mg of the resultant compounds of    Example 17A and 0.2 mL of diisopropylethylamine in 10 mL of    dichloromethane was treated with 256 mg of di-tert-butyl    dicarbonate. After 24 hours, the mixture was diluted with    dichloromethane. The mixture was washed with water, 5% NaHCO3, 0.5 N    HCl, brine then dried over MgSO₄, filtered and concentrated in    vacuo. The residue was purified by low pressure silica gel column    chromatography using a 20% ethyl acetate/dichloromethane as eluent    to yield 154.6 mg of the fast moving diastereomer A, later    identified as having the anti configuration at the hydroxyl center;    98.8 mg of the slower moving diastereomer B, having the syn    configuration at the hydroxyl center, and 204.6 mg of the mixed    diastereomers A and B. TLC: Rf=0.60, 0.67, 40% EtOAc/CH₂Cl₂.-   C. Compound 17. A solution of 64.6 mg of the resultant compounds of    Example 17B, diastereomer B, in 1.5 mL of dichloromethane was    treated with 1.5 mL of trifluoroacetic acid. After 4 hours, the    mixture was concentrated in vacuo to yield the amine    trifluoroacetate salt. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solution    of 17.8 mg of the resultant trifluoroacetate salt in 1 mL of    dichloromethane was sequentially added 0.3 mL of saturated NaHCO₃, a    small amount of solid NaHCO₃ and 10.7 mg of    4-acetamido-benzenesulphonyl chloride. After 3 hours, the mixture    was diluted with dichloromethane. The two layers were separated and    the aqueous layer was extracted once with dichloromethane. The    combined organic layer was washed with brine then dried over MgSO₄,    filtered and concentrated in vacuo. The residue was purified by    preparative HPLC to yield 14.4 mg of the title compound as a white    solid; TLC: Rf=0.54, 10% CH₃OH/CH₂Cl₂; HPLC, Rt=13.58 min; (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 18

Compound 18. To a solution of 20.8 mg (0.041 mmol) of the crudetrifluoroacetate salt obtained as from Example 17B, diastereomer B, in 1mL of dichloromethane was sequentially added 0.3 mL of saturated NaHCO₃,a small amount of solid NaHCO₃ and 13.6 mg (0.054 mmol) of2-acetamido-4-methyl-5-thiazolesulphonyl chloride. After 3 hours, themixture was diluted with dichloromethane. The two layers were separatedand the aqueous layer was extracted once with dichloromethane. Thecombined organic layer was washed with brine then dried over MgSO₄,filtered and concentrated in vacuo. The residue was purified bypreparative HPLC to yield 4.8 mg of the title compound as a white solid;TLC: Rf=0.50, 10% CH₃OH/CH₂Cl₂; HPLC: Rt=13.35 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 19

-   A. Sodium 3-acetamidobenzenesulfonate. A solution of 118.6 mg (0.55    mmol) of 3-acetamidobenzenesulfonic acid in 0.5 mL of water was    treated with 0.55 mL (0.55 mmol) of 1.0 N NaOH at 0° C. After    stirring at room temperature for 4 hours, the mixture was    concentrated to dryness and used without subsequent purification.-   B. 3-Acetamidobenzenesulfonyl chloride. The crude mixture from    Example 19A was cooled to 0° C. and 0.29 g (1.38 mmol) of phosphorus    pentachloride was added. The mixture of solid was stirred for 3    hours then 5 mL dichloromethane was added. After 24 hours, the    slurry was filtered and concentrated in vacuo to yield 81.4 mg of    solid product which was used without subsequent purification. TLC:    Rf=0.50, 40% EtOAC/CH₂Cl₂.-   C. Compound 19. A solution of 82.7 mg (0.098 mmol) of diastereomer    B, obtained in Example 17B, in 2 mL of dichloromethane was treated    with 2 mL of trifluoroacetic acid. After 4 hours, the mixture was    concentrated in vacuo to yield the amine trifluoroacetate salt which    was used without further purification; TLC: Rf=0.11, 10%    CH₃OH/CH₂Cl₂. A solution of this salt (entire yield) in 2 mL of    dichloromethane was treated sequentially with 0.5 mL of saturated    NaHCHO₃, small amount of solid NaHCO₃ and a solution of 81.4 mg    (0.046 mmol) of the resultant compound of Example 19B. After 3    hours, the mixture was diluted with dichloromethane. The two layers    were separated and the aqueous layer was extracted once with    dichloromethane. The combined organic layer was washed with brine    then dried over MgSO₄, filtered and concentrated in vacuo. The    residue was purified by preparative HPLC to yield 24.7 mg of the    title compound as a white solid; TLC: Rf=0.42, 10% CH₃OH/CH₂Cl₂;    HPLC: Rt=13.8 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 20

Compound 20. A solution of 209.0 mg (0.24 mmol) of the resultantcompound of Example 17B, diastereomer B, in 5 mL of dichloromethane wastreated with 5 mL of trifluoroacetic acid. After 4 hours, the mixturewas concentrated in vacuo. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solutionof this residue in 2 mL of dichloromethane was sequentially added 0.5 mLof saturated NaHCO₃, a small amount of solid NaHCO₃ and 70.2 mg (0.32mmol) of benzofurazan-4-sulphonyl chloride. After 3 hours, the mixturewas diluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 108.0 mg of the title compound as a white solid; TLC: Rf=0.60, 10%CH₃OH/CH₂Cl₂; HPLC: Rt=14.95 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 21

Compound 21. The resulting compound of Example 17B, diastereomer B, (228mg, 0.27 mmol) was dissolved in 1:1 CH₂Cl₂/TFA (10 mL), and the reactionmixture stirred for 3.5 hours, then concentrated to dryness to affordthe product trifluoroacetate salt as a yellow solid which was used inthe next reaction without purification. To a solution of this residue(34.7 mg, 0.05 mmol) in CH₂Cl₂ (3 mL) was added Heunig's base (41 μl,0.24 mmol) and dimethylsulfamoyl chloride (11 μl, 0.09 mmol), and thereaction was stirred for 17 hours at room temperature. The reactionmixture was then diluted with CH₂Cl₂ and washed with saturated NH₄Cl,and the organic layer was dried over MgSO₄. Filtration and concentrationprovided a residue which was chromatographed on a silica gel columnusing 8% CH₃OH/CH₂Cl₂ as eluent, yielding the desired compound which wasfurther subject to purification by preparative HPLC. HPLC: Rt=13.8minutes. TLC: Rf=0.40, 8% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 22

-   A. N^(α)-isocyano-L-valine methyl ester. To the HCl salt of valine    methyl ester (2.08 g, 12.40 mmol) in toluene (20 mL) was added a 20%    solution of phosgene in toluene (32 mL, 62.00 mmol), and the    solution was heated at reflux for 12 hours. The reaction was then    cooled to room temperature and concentrated in-vacuo to give a pale    yellow liquid which was used in the subsequent reaction without    purification. TLC: Rf=0.88, 50% Hexane/EtOAc; (¹H)-NMR (CDCl₃)    consistent with structure.-   B. N^(α)-(2-pyridylmethyl)-oxycarbonyl-L-valine methyl ester. A    mixture of 2-pyridylcarbinol (941 μl, 9.75 mmol) and the resulting    compound of Example 22A (1.28 g, 8.12 mmol) were allowed to stir in    CH₂Cl₂ (7 mL) for 12 hours, then the reaction was concentrated and    the residue chromatographed with 50% hexane/EtOAc to afford 2.03    grams of the title compound as a colorless oil. TLC: Rf=0.26, 50%    Hexane/EtOAc; (¹H)-NMR (CDCl₃) consistent with structure.-   C. N^(α)-(2-pyridylmethyl)-oxycarbonyl-L-valine. A solution of the    resulting compound of Example 22B (634 mg, 2.38 mmol) in a 1/1    mixture of 1N HCl/THF (6 mL) containing 12 N HCl (0.5 mL) was    allowed to stir at room temperature over 15 hours, but much starting    material was still present by TLC. Hence, more 12 N HCL was added (1    mL), and the reaction stirred an additional 48 hours. The reaction    was then concentrated to dryness and diluted with CH₂Cl₂, yielding    the desired carboxylic acid as an insoluble resin which was washed    with additional CH₂Cl₂, providing 22C which contained minor    quantities of 22B. This material was used in the subsequent reaction    without further purification. TLC: Rf=0.11, 8% CH₃OH/CH₂Cl₂;    (¹H)-NMR (CDCl₃) consistent with structure.-   D. Compound XXX (P=(2-pyridylmethyl)-oxycarbonyl, R³=isopropyl,    R^(3′)=H, D′=isobutyl, P′=tert-butoxycarbonyl). To the resulting    compound of Example 21B (277 mg, 0.82 mmol) in CH₂Cl₂ (5 mL) was    added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride    (210 mg, 1.10 mmol), the acid 22C (402 mg, 1.10 mmol), and    1-hydroxybenzotriazole hydrate (148 mg, 1.10 mmol). The reaction    proceeded for 12 hours at room temperature, then was diluted with    CH₂Cl₂ and washed successively with saturated NH₄Cl and NaHCO₃, and    the organic layer was dried over MgSO₄. Filtration and concentration    provided a residue which was chromatographed on a silica gel column    using 17% THF/CH₂Cl₂ as eluent, yielding 396 mg of product. TLC:    Rf=0.26, 17% THF/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.-   E. Compound 22. The resulting compound of Example 22D (396 mg, 0.69    mmol) was dissolved in 90% aqueous TFA (11 mL), and the reaction    mixture stirred for 3 hours at room temperature, then was    concentrated to dryness. To a solution of this residue (231 mg. 0.33    mmol) in CH₂Cl₂ (5 mL) was added excess solid NaHCO₃ (approx. 1    gram) and saturated aqueous NaHCO₃ (20 μl), followed by    N-acetylsulfanilyl chloride (116 mg, 0.50 mmol), and the reaction    proceeded for 12 hours at room temperature. The reaction mixture was    then diluted with CH₂Cl₂ and washed with saturated NaHCO₃, and the    organic layer was dried over MgSO₄. Filtration and concentration    provided a residue which was chromatographed on a silica gel column    using 8% CH₃OH/CH₂Cl₂ as eluent, yielding the desired compound which    was further subject to purification by preparative HPLC (76.1 mg of    3 was obtained). HPLC: Rt=12.1 minutes. TLC: Rf=0.46, 8%    CH₃OH/CH₂Cl₂; NMR (CDCl₃): 8.76 (d, 1H), 8.40 (br s, 1H), 8.26 (t,    1H), 7.72 (d, 2H), 7.67 (d, 2H), 7.58 (d, 2H), 7.37 (d, 1H), 7.25    (m, 4H), 7.16 (br d, 1H), 6.47 (d, 1H), 5.65 (d, 1H), 5.26 (d, 1H),    4.32 (m, 1H), 3.91 (t, 1H), 3.83 (m, 1H), 3.23 (d, 1H), 3.05 (m,    2H), 2.68-3.10 (m, 3H), 2.22 (m, 3H), 2.0 (m, 1H), 1.82 (m, 1H),    0.85 (d, 3H), 0.80 (d, 3H), 0.71 (d,3H), 0.65 (d, 3H).

EXAMPLE 23

Compound 23. Prepared by the same route as described for Example 22,except 4-pyridylcarbinol was utilized for reaction with the product ofExample 22A. HPLC: Rt=12.0 minutes. TLC: Rf=0.50 (8% CH₃OH/CH₂Cl₂);(1H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 24

Compound 24. A solution of the resulting compound of the trifluoroaceticacid deprotection of Example 22D (as described in Example 22E; 215 mg,0.31 mmol) in CH₂Cl₂ at room temperature was treated withdiisopropylethylamine (214 μl, 1.23 mmol) and dimethylsulfamoyl chloride(40 μl, 0.37 mmol) in CH₂Cl₂ at room temperature in CH₂Cl₂ at roomtemperature for 12 hours. The reaction mixture was concentrated andchromatographed on a silica gel column with 5% CH₃OH/CH₂Cl₂ as eluent,yielding the desired compound which was further subject to purificationby preparative HPLC (9.5 mg obtained). HPLC: Rt=14.4 minutes. TLC:Rf=0.88, 11% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 25

Compound 25. This compound was prepared by the route described forExample 22, except that 3-pyridylcarbinol was utilized for reaction withthe compound produced in Example 22A, and in the reaction correspondingto 22E, the trifluoracetate-deprotected material was reacted withbenzofurazan-4-sulphonyl chloride. HPLC: Rt=9.4 minutes. TLC: Rf=0.10,11% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 26

Compound 26. A solution of the resulting compound from thetrifluoroacetic acid deprotection of Example 22D (as described inExample 22E; 27 mg, 0.14 mmol) in CH₂Cl₂ was treated with excess solidNaHCO₃ (approx. 1 gram) and saturated aqueous NaHCO₃ (7 μl), thenstirred vigorously at room temperature for 3 hours. The reaction mixturewas decanted from the solids, concentrated, then the residue waspurified directly by preparation HPLC (3.0 mg of white solid obtained).HPLC: Rt=14.7 minutes; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 27

Compound 27. A solution of 33 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 20 mg of N,N-diisopropylethylamine and 9.3mg of allyl chloroformate. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂C₂):diethyl ether to yield 24 mg of the titlecompound as a white solid. TLC: Rf=0.53, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=14.53 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 28

Compound 28. A solution of 47.5 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 28.7 mg of N,N-diisopropylethylamine and15.2 mg of isobutyl chloroformate. The mixture was stirred 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 45 mg of the titlecompound as a white solid. TLC: Rf=0.60, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=15.58 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 29

Compound 29. A solution of 35.6 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 21.5 mg of N,N-diisopropylethylamine and0.083 nL of 1.0 M isopropyl chloroformate. The mixture was stirred 3 hand then concentrated in vacuo. The residue was taken up in ethylacetate and washed with 0.5 N HCl and saturated NaCl then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of5:10:85 NH₄OH/CH₃OH/CH₂Cl₂:diethyl ether to yield 33.2 mg of the titlecompound as a white solid. TLC: Rf=0.56, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=14.81 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 30

-   A. (2-Pyrrolidinonyl-hydroxyethyl-N-hydroxysuccinimdyl carbonate. A    solution of 572 mg of 1-(2-hydroxyethyl)-2-pyrrolidinone and 1.70 g    of N,N′-disuccinimidyl carbonate in acetonitrile was treated, at    ambient temperature under an atmosphere of nitrogen, with 1717 mg of    N,N-diisopropylethylamine. The mixture was stirred for 14 h and    concentrated in vacuo. The residue was taken up in ethyl acetate and    washed with saturated NaHCO₃, saturated NaCl, then dried over MgSO₄,    filtered, and concentrated in vacuo to yield 200 mg of a white    solid. TLC: Rf=0.56, 10% isopropanol in CH₂Cl₂; (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound 30. A solution of 68 mg of the resultant compound of    Example 30A in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 32 mg of the resultant    compound of Example 40A and 39 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 4 h, diluted with CH₂Cl₂, washed    with saturated NaHCO₃ and saturated NaCl, then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was subjected to    preparative thin layer silica gel chromatography using a 2:1 mixture    of 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂:diethyl ether to yield 45 mg of    residue. About 20 mg of this residue was purified by preparative    HPLC to yield 13.5 mg of the title compound as a white solid. TLC:    Rf=0.47, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂. HPLC: Rt=12.79 mid; (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 31

Compound 31. A solution of 39.7 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 24 mg of N,N-diisopropylethylamine and 14.5mg of phenyl chloroformate. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of5:10:85 NH₄OH/CH₃OH/CH₂Cl₂:diethyl ether to yield 39.7 mg of the titlecompound. TLC: Rf=0.53, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂. HPLC: Rt=15.22 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 32

Compound 32. A solution of 391 mg of the resultant compound of Example39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 271 mg of4-fluorobenzenesulfonyl chloride and 117 mg of sodium bicarbonate. Themixture was stirred for 14 h, diluted with CH₂Cl₂, washed with saturatedNaCl then dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was purified by low pressure silica gel chromatography using 5%diethyl ether in CH₂Cl₂ as eluent to yield 420 mg of the title compoundas a white solid. TLC: Rf=0.20, 5% diethyl ether in CH₂Cl₂. HPLC:Rt=17.41 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 33

Compound 33. A solution of 30 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 18.1 mg of N,N-diisopropylethylamine and9.3 mg of benzyl isocyanate. The mixture was stirred 14 h and thenconcetrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a mixture of5:10:85 NH₄OH/CH₃OH/CH₂Cl₂ to yield 30.2 mg of the title compound as awhite solid. TLC: Rf=0.56, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂. HPLC: Rt=14.36min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 34

Compound 34. A solution of 55 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 33.3 mg of N,N-diisopropylethylamine and17.8 mg of 2-methoxyethyl chloroformate. The mixture was stirred for 3 hand then concentrated in vacuo. The residue was taken up in ethylacetate and washed with 0.5 N HCl and saturated NaCl then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 48.1 mg of the titlecompound as a white solid. TLC: Rf=0.56, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=13.43 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 35

-   A. Compound XXI (D′=isobutyl, A′=4-fluorophenyl, hydrochloride    salt). A solution of 398 mg of the resultant compound of Example 32    in ethyl acetate was treated at −20° C. with HCl gas. The HCl was    bubbled through the mixture for 20 min over which time the    temperature was allowed to warm to 20° C. Nitrogen was then bubbled    through the mixture for 15 min and solvent removed in vacuo to yield    347 mg of the title compound as a white solid. TLC: Rf=0.82, 5:10:85    NH₄OH/CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound 35. A solution of 111 mg of the resultant compound of    Example 35A in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 118 mg of the resultant    compound of Example 48A and 133 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 14 h, diluted with CH₂Cl₂,    washed with saturated NaHCO₃ and saturated NaCl, then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was    subjected to preparative thin layer silica gel chromatography using    5% CH₃OH in CH₂Cl₂ to yield 98.8 mg of the title compound as a white    solid. TLC: Rf=0.48, 5% CH₃OH in CH₂Cl₂. HPLC: Rt=15.18 min;    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 36

Compound 36. A solution of 48 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 29.0 mg of N,N-diisopropylethylamine and15.1 mg of 3-butenyl chloroformate. The mixture was stirred for 3 h andthen concentrated in vacuo. The residue was taken up in ethyl acetateand washed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 43.8 mg of the titlecompound as a white solid. TLC: Rf=0.83, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂;Rf=0.24, 5% diethyl ether in CH₂Cl₂. HPLC: Rt=14.76 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 37

Compound 37. A solution of 99 mg of the resultant compound of Example51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 83.2 mg of3,4-dichlorobenzenesulfonyl chloride and 29 mg of sodium bicarbonate.The mixture was stirred for 14 h, diluted with CH₂Cl₂, washed withsaturated NaCl then dried over MgSO₄, filtered, and concentrated invacuo. The residue was subjected to operative thin layer silica gelchromatography using 5% CH₃OH in CH₂Cl₂ to yield 107 mg of the titlecompound as a white solid. TLC: Rf=0.35 (5% CH₃OH in CH₂Cl₂). HPLC:Rt=17.27 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 38

Compound 38. To a solution of 32 mg of the resultant compound of Example35A in CH₂Cl₂ was added, at ambient temperature under an atmosphere ofnitrogen, 14 mg of benzyl chloroformate and 21 mgN,N-diisopropylethylamine. The mixture was stirred for 4 h, diluted withCH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl, then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using 10% diethyl etherin CH₂Cl₂ as eluent to yield 33 mg of product. TLC: Rf=0.62, 10% diethylether in CH₂Cl₂. HPLC: Rt=17.27 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 39

-   A. Compound XXI (D′=isobutyl, P=tert-butoxy carbonyl, P′=H). A    solution of 4.1 g of epoxide XX (P=Boc) in 30 mL of ethanol was    treated with 22.4 mL of isobutylamine and heated under reflux for    1 h. The mixture was concentrated to yield the title compound as a    white solid which was used without subsequent purification. NMR    (CDCl₃): δ0.91 (d, 3H); 0.93 (d, 3H); 1.37 (s, 9H); 1.68 (br s, 2H);    2.40 (d, 2H); 2.68 (d, 2H); 2.87 (dd, 1H); 2.99 (dd, 1H); 3.46 (dd,    1H); 3.75 (br s, 1H); 3.80 (br s, 1H); 4.69 (d, 1H); 7.19-7.32 (m,    4H).-   B. Compound 39. To a solution of 514.1 mg of the resultant compound    of Example 39A in dichloromethane (10 mL) was added aqueous sodium    bicarbonate (5 mL) and N-acetylsulfanilyl chloride (428.4 mg). After    14 h, the resulting mixture was diluted with ethyl acetate, washed    with sodium bicarbonate, saturated brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by low pressure silica gel column chromatography using 20%    ethyl acetate in dichloromethane eluent to yield 714.4 mg of the    title product. TLC: Rf=0.63, 60% ethyl acetate/dichloromethane,    HPLC: Rt=15.3 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 40

-   A. Compound XXII (D′=isobutyl, P=H, E=4-acetamidophenyl),    hydrochloride salt. To a solution of 691.4 mg (1.296 mmol) of the    resultant compound of Example 39B in ethyl acetate (20 mL) at    −20° C. was bubbled anhydrous HCl gas for 10 min. The ice bath was    removed and after an additional 15 min., the reaction mixture was    sparged with nitrogen then concentrated in vacuo to provide 610 mg    of title product which was used without subsequent purification.-   B. Compound 40. A solution of 41.5 mg of the resultant crude    compound of Example 40A in 5 mL of dichloromethane was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 18.1 mg of L-dihydroorotic acid, 0.031 mL (0.176    mmol) diisopropylethylamine, 15.5 mg (0.115 mmol) of    1-hydroxybenzotriazole hydrate, 22 mg (0.115 mmol) EDC. After 1 h,    the slurry was treated with 1 mL of dimethylformamide. The mixture    was stirred for 16 h and then concentrated in vacuo. The residue was    taken up in ethyl acetate and washed with water and saturated brine,    dried over magnesium sulfate, filtered and concentrated in vacuo.    The residue was purified by thin layer preparative chromatography    using (1/2/17 v/v/v/30% ammonium hydroxide/methanol/dichlomethane)    eluent to provide 34.2 mg of the title product. TLC: Rf=0.33, 1/2/17    v/v/v/30% ammonium hydroxide/methanol/dichlomethane). HPLC: Rt=11.3    min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 41

Compound 41. To a solution of 42.8 mg of the resultant compound ofExample 40A in 5 mL dichloromethane was added sequentially, at ambienttemperature under an atmosphere of nitrogen., 17.2 mg of N-tert-butylglyoxalic acid, 0.032 mL diisopropylethylamine, 16 mg of1-hydroxybenzotriazole hydrate, 22.6 mg EDC. The mixture was stirred.for 16 h and then concentrated in vacuo. The residue was taken up inethyl acetate and washed with water, 0.5 N hydrochloric acid, washedwith sodium bicarbonate, saturated brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by thinlayer preparative chromatography using 40% ethyl acetate/dichloromethaneeluent to provide 14.9 mg of the title product. TLC: Rf=0.47, 40% ethylacetate/dichloromethane, HPLC: Rt=15.2 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 42

Compound 42. To a solution of 43.5 mg of the resultant crude compound ofExample 40A in 5 mL dichloromethane was added sequentially at ambienttemperature, under an atmosphere of nitrogen, 13.0 mg of succinamicacid, 0.024 mL diisopropylethylamine, 15.0 mg of 1-hydroxybenzotriazolehydrate, and 21.3 mg EDC. The mixture was stirred for 16 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with sodium bicarbonate, saturated brine, dried over magnesiumsulfate, filtered and concentrated in vacuo. The residue was purified bythin layer preparative chromatography using (1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane) eluent to provide 35.3 mg of the titleproduct. TLC: Rf=0.25, 1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane, HPLC: Rt=11.6 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 43

Compound 43. To a solution of 42.8 mg of the resultant compound ofExample 40A in 5 mL dichloromethane was added sequentially, at ambienttemperature under an atmosphere of nitrogen, with 14.1 mg ofL-pyroglutamic acid, 0.024 mL diisopropylethylamine, 14.8 mg of1-hydroxybenzotriazole hydrate, 20.9 mg EDC. The mixture was stirred for16 h and then concentrated in vacuo. The residue was taken up in ethylacetate and washed with water, 0.5 N hydrochloric acid, washed withsodium bicarbonate, saturated brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by thinlayer preparative chromatography using (1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane) eluent to provide 29.9 mg of the titleproduct. TLC: Rf=0.33, 1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane, HPLC: Rt=11.7 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 44

-   A. 3-Pyridylmethyl-N-hydroxysucchinimdyl carbonate. To a solution of    181.0 mg of 3 pyidinecarbinol in 5 mL acetonitrile was added    sequentially at ambient temperature under an atmosphere of nitrogen,    with 0.72 mL diisopropylethylamine and 354.1 mg of    N,N′-disuccinimidyl carbonate. After 4 h, the resultant mixture was    concentrated in vacuo to provide a yellow solid which was used    without subsequent purification.-   B. Compound 44. To a solution of 58.1 mg of the resultant crude    compound of Example 40A in 3 mL of dichloromethane was added    sequentially, at ambient temperature under an atmosphere of    nitrogen, 0.075 mL diisopropylethylamine and 46.3 mg of the    resultant compound of Example 20A. The mixture was stirred for 16 h    and then concentrated in vacuo. The residue was taken up in diethyl    ether and extracted into 3×25 mL of 0.5N HCl. The combined aqueous    extracts were adjusted to pH 8 with solid sodium bicarbonate and    extracted into 3×25 mL ethyl acetate. The combined organic extracts    were washed with saturated brine, dried over magnesium sulfate,    filtered, and concentrated in vacuo. The residue was purified by    thin layer preparative chromatography using (1/2/17/20 v/v/v/30%    ammonium hydroxide/methanol/dichlomethane/diethyl ether) eluent to    provide 10.3 mg of the title product. TLC: Rf=0.4, 1/2/17/20    v/v/v/30% ammonium hydroxide/methanol/dichlomethane/diethyl ether,    HPLC: Rt=11.8 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 45

Compound 45. To a solution of 28.3 mg of the resultant compound ofExample 39A in 4 mL of dichloromethane was added 1 mL saturated aqueoussodium bicarbonate solution, 9.2 mg sodium bicarbonate, and 0.013 mL ofbenzenesulfonyl chloride. After 14 h, the resulting mixture was dilutedwith ethyl acetate, washed with saturated brine, dried over magnesiumsulfate, filtered and concentrated in vacuo. The residue was purified bythin layer preparative chromatography using 10% diethylether/dichloromethane eluent to provide 19.3 mg of the title product.TLC: Rf=0.84, 25% diethyl ether/dichlormethane, HPLC: Rt=17.2 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 46

Compound 46. To a solution of 47.0 mg (0.140 mmol) of the resultantcompound of Example 39A in 4 mL of dichloromethane was added 1 mLsaturated aqueous sodium bicarbonate solution, 17.6 mg of solid sodiumbicarbonate, and 41.4 mg of 2,4 dimethylthiazole-5-sulfonyl chloride.After 14 h, the resulting mixture was diluted with ethyl acetate, washedwith saturated brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by thin layerpreparative chromatography using 25% ethyl acetate/dichloromethaneeluent to provide 34.6 mg of the title product. TLC: Rf=0.44, 25%diethyl ether/dichloromethane, HPLC: Rt=16.4 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 47

Compound 47. To a solution of 50.7 mg of the resultant compound ofExample 39A in 4 mL of dichloromethane was added 1 mL saturated aqueoussodium bicarbonate solution, 15.2 mg of solid sodium bicarbonate, and2-fluorobenzenesulfonyl chloride 35.2 mg. After 14 h, the resultingmixture was diluted with ethyl acetate, washed with saturated brine,dried over magnesium sulfate, filtered and concentrated in vacuo. Theresidue was purified by thin layer preparative chromatography using 10%diethyl ether/dichloromethane eluent to provide 40.5 mg of the titleproduct. TLC: Rf=0.44, 25% diethyl ether/dichloromethane, HPLC: Rt=17.2min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 48

-   A. N-succinimidlyl-(S)-3-tetrahydrofuryl carbonate. To a solution of    12.5 mL of 1.93 M phosgene in toluene at 0-5° C. was added 1.3 g of    (S)-(+)-3-hydroxy-tetrahydrofuran. After stirring for 2 h, the    reaction mixture was sparged with nitrogen and then concentrated to    dryness in vacuo to provide 1.486 g of crude chloroformate. This    material was taken up in 10 mL of acetonitrile and treated    sequentially at ambient temperature under an atmosphere of nitrogen    with 1.17 g of N-hydroxysuccinimide and 1.41 mL of triethylamine.    After stirring for 14 h, the reaction mixture was concentrated in    vacuo to provide 3.44 g of the title product as a white solid.-   B. Compound 48. To a solution of 87.2 mg of the resultant compound    of Example 40A in 5 mL of dichloromethane was added sequentially, at    ambient temperature under an atmosphere of nitrogen, 0.113 mL    diisopropylethylamine and 68 mg of the resultant compound of Example    48A. The mixture was stirred for 16 h and then concentrated in    vacuo. The residue was taken up in ethyl acetate and washed with    water, 0.5 N HCl, saturated sodium bicarbonate, saturated brine,    dried over magnesium sulfate, filtered, and concentrated in vacuo.    The residue was purified by silica gel chromatography using    (3/6/20/65 v/v/v/v/30% ammonium hydroxide/methanol/diethyl    ether/dichlomethane) eluent followed by crystallization from a    mixture of dichloromethane, diethyl ether, and hexanes to provide 58    mg of the title product. TLC: Rf=0.17, 75% ethyl    acetate/dichloromethane, HPLC: Rt=13.1 min.; (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 49

Compound 49. Following the procedure described in Example 83, a solutionof the resultant compound of Example 39A in CH₂Cl₂ is reacted with2,4-difluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄ filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography usingan appropriate solvent system to yield the title product.

EXAMPLE 50

Compound 50. A solution of 30 mg of the resulting compound of Example 58and 9 μL of dimethysulfamoyl chloride in 10 mL of CH₂Cl₂ was reacted inthe same manner as described for Example 14. After workup andpurification by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 6.5 mg of thetitle compound was obtained. TLC: Rf=0.2, 3% CH₃OH in CH₂Cl₂. HPLC:Rt=15.96 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 51

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=isobutyl,    A′=benzyloxycarbonyl). To a solution of the resultant compound of    Example 39A (2.5 g, 7.43 mmol) in CH₂Cl₂ (50 mL) was added    triethylamine (2.1 mL, 14.9 mmol) followed by addition of benzyl    chloroformate (1.2 mL, 8.1 mmol). The mixture was allowed to stir at    ambient temperature for 6 h. The solution was diluted with 1 L of    CH₂Cl₂ and washed with water. The organics were dried over anhydrous    MgSO₄, concentrated under reduced pressure, then purified via silica    gel chromatography. Gradient solvent system: CH₂Cl₂ followed by 3:97    methanol/CH₂Cl₂. The title compound (2.97 g, was obtained as a    colorless oil. TLC: Rf=0.14, 3:97 methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXI (A=H, D′=isobutyl, A′=benzyloxycarbonyl,    hydrochloride salt). To a solution of 1.5 g (3.187 mmol) of the    resultant compound of Example 51A in ethyl acetate (25 mL) at    −20° C. was bubbled anhydrous HCl gas for 10 min. The ice bath was    removed and after an additional 15 min. the reaction mixture was    sparged with nitrogen, then concentrated in vacuo to provide 1.29 g    of title product as a white solid which was used directly for    ensuing reaction. TLC: Rf=0.14, 10% methanol/CH₂Cl₂.-   C. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl, D′=isobutyl,    A′=benzyloxycarbonyl). To a solution of 1.077 g of the resultant    crude compound of Example 51B (2.647 mmol) in acetonitrile (10 mL)    was added sequentially at ambient temperature under an atmosphere of    nitrogen, 1.61 mL (9.263 mmol) of diisopropylethylamine and 910 mg    (3.97 mmol) of the resultant compound of Example 48A. After stirring    for 3 h, an additional 223 mg (0.973 mmol) of the resultant compound    of Example 48A was added. The mixture was stirred for 16 h and then    concentrated in vacuo. The residue was taken up in ethyl acetate and    washed with water, 0.5 N HCl, saturated sodium bicarbonate,    saturated brine, dried over magnesium sulfate, filtered, and    concentrated in vacuo. The residue was purified by low pressure    silica gel column chromatography using a gradient 10% to 25% ethyl    acetate in CH₂Cl₂ eluent to yield 1.025 g of the title product as a    white solid. TLC: Rf=0.10, 10% ethyl acetate/CH₂Cl₂; (¹H)-NMR    (CDCl₃) consistent with structure.-   D. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl, D′=isobutyl,    A′=H). A solution of 872 mg (1.799 mmol) of the resultant compounds    of Example 51C in (10 mL) of ethyl alcohol was added, at ambient    temperature under a nitrogen atmosphere, to a slurry of 87 mg (10%    by weight) of 10% palladium on carbon in (5 mL) ethyl alcohol and    hydrogenated for 16 h under a slight positive pressure of hydrogen.    The mixture was filtered and concentrated in vacuo to yield 553.2 mg    of the title product as a colorless glass which was used directly    for ensuing reaction. TLC: Rf=0.46, 10% methanol/CH₂Cl₂.-   E. Compound 51. To a solution of 72.7 mg (0.207 mmol) of the    resultant compound of Example 51D in CH₂Cl₂ (4 mL) was added aqueous    sodium bicarbonate (1 mL), solid sodium bicarbonate 22.6 mg (0.27    mmol), and 2-(pyrid-2-yl)-thiophene-5-sulfonyl chloride 64.6 mg,    (0.249 mmol). After 14 h, the resulting mixture was diluted with    ethyl acetate, washed with saturated brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by thin layer preparative chromatography using 15 to 30%    ethyl acetate/CH₂Cl₂ eluent to provide 53 mg of the title product as    a white solid. TLC; RF=0.25, 25% ethyl acetate/CH₂Cl₂, HPLC: Rt=15.3    min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 52

-   A. N-hydroxysuccinimidyl-(RS)-3-hydroxyl-tetrahydrofuryl carbonate.    The title compound was prepared as described in Example 48A starting    with 1.0 g of (RS)-3-hydroxy-tetrahydrofuran and yielding 2.33 g of    a white solid.-   B. Compound 52. To a solution of 105 mg of the resultant compound of    Example 35A in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, 112 mg of the resultant compound of Example    52A and 126 mg N,N-diisopropylethylamine. The mixture was stirred    for 4 h, diluted with CH₂Cl₂, washed and saturated NaHCO₃ and    saturated NaCl, then dried over MgSO₄, filtered, and concentrated in    vacuo. The residue was purified by low pressure silica gel    chromatography using 5% CH₃OH in CH₂Cl₂ as eluent to yield 101.4 mg    of product. TLC: Rf=0.52, 5% CH₃OH in CH₂Cl₂. HPLC: Rt=15.05 min.    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 53

Compound 53. To a solution of 72.3 mg (0.19 mmol) of the resultantcompound of Example 51D in CH₂Cl₂ (4 mL) was added aqueous sodiumbicarbonate (1 mL), solid sodium bicarbonate 19.2 mg (0.228 mmol), and4-acetamido-3-chlorobenzene sulfonyl chloride 61.1 mg, (0.228 mmol).After 14 h, the resulting mixture was diluted with EtOAc, washed withsaturated brine, dried over magnesium sulfate, filtered and concentratedin vacuo. The residue was purified by low pressure silica gel columnchromatography using 20% to 45% EtOAc/CH₂Cl₂ eluent to provide 49.1 mgof the title product. TLC: RF=0.29, 50% EtOAc/CH₂Cl₂, HPLC: Rt=13.9 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 54

Compound 54. A solution of 260 mg of the resulting Compound of 39A and45 mg of 3-acetamido-4-fluorobenzenesulfonyl chloride in 10 mL of CH₂Cl₂was reacted in the same manner as described for Example 14. After workupand purification by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 1.4 mg of thetitle compound was obtained. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC:Rt=15.63 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 55

Compound 55. 35.0 mg of the resulting compound of Example 54 was treatedwith 1 mL of 90% aqueous TFA and allowed to stand for 12 h. The mixturewas concentrated in vacuo and the residue taken up in 10 mL of dryCH₂Cl₂, treated with 34 μL of DIEA (0.23 mmoles) and 20 mg of1-benzyl-3-tert-butyl-1H-pyrazole-5-carbonyl chloride. The mixture wasstirred for 1.5 h, then diluted with in CH₂Cl₂, and washed with 1 N HCl.After drying over MgSO₄ and concentrating in vacuo, a portion of themixture was purified by preparative reversed-phase C₁₈ HPLC using alinear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA for elution toobtain 1.1 mg of the title compound. TLC: Rf=0.8, 5% CH₃OH in CH₂Cl₂.HPLC: Rt=18.25 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 56

-   A. S(−)-1-phenylethyl-N-hydroxysuccinimdyl carbonate. The title    compound was prepared from 9.5 μL of S(−)-1-phenylethanol and 30 m    of N,N-disuccinimidyl carbonate as described in Example 44A. The    resulting material was used without subsequent purification;    (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound 56. 45.0 mg Of the resulting compound of Example 58 was    treated with 1 mL of 90% aqueous TFA and allowed to stand for 12 h.    The mixture was concentrated in vacuo and the residue taken up in 15    mL of dry CH₂Cl₂, treated with the above mixed anhydride and 65 μL    of triethylamine. The mixture was stirred for 14 h then diluted with    ethyl acetate and washed with saturated sodium bicarbonate solution    and saturated brine, dried over magnesium sulfate, filtered and    concentrated in vacuo. A portion of the mixture was purified by    preparative reversed-phase C₁₈ HPLC using a linear gradient of    mixture was purified by preparative reversed-phase C₁₈ HPLC using a    linear gradient of 35% of 100% CH₃CN/H₂O with 0.1% TFA for elution    to obtain 1.1 mg of the title compound. TLC: Rf=0.5, 3% CH₃OH in    CH₂Cl₂. HPLC: Rt=17.44 min; (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 57

Compound 57. 30 mg of the resultant compound of Example 58 was treatedwith 1 mL of 90% aqueous TFA and allowed to stand for 12 h. The mixturewas concentrated in vacuo and the residue taken up in 25 mL of dryCH₂Cl₂ washed and saturated sodium bicarbonate solution, dried overmagnesium sulfate, filtered and concentrated in vacuo. A solution of 14mg of the resultant free amine in 10 mL of CH₂Cl₂ was treated with 6 μLof phenoxyacetyl chloride and 12 μL of triethylamine. The mixture wasstirred under an inert atmosphere for 1 h, then diluted in CH₂Cl₂ andwashed with 1 N HCl. After drying over MgSO₄ and concentrating in vacuo.A portion of the mixture was purified by preparative reversed-phase C₁₈HPLC using a linear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA aseluant to obtain 16.5 mg of the title compound. TLC: Rf=0.25, 3% MeOH inCH₂Cl₂. HPLC: Rt=16.6 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 58

Compound 58. A solution of 500 mg of the resulting compound of Example39A and 370 mg of benzofurazan-4-sulfonyl chloride in 10 mL of CH₂Cl₂was reacted in the same manner as described for Example 14. Afterworkup, the title compound was obtained by crystallization from hotethanol. Further purification of this material by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluent gave 2.0 mg of the title compound. TLC: Rf=0.35,3% CH₃OH in CH₂Cl₂. HPLC: Rt=17.00 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 59

-   A. R(+)-1-phenylethyl-N-hydroxysuccinimdyl carbonate. The title    compound was prepared from R(+)-1-phenylethanol as described in    Example 56A to yield a white solid. The resulting material was used    directly for subsequent reaction; (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound 59. A 36 mg portion of the resultant compound of Example    58 and 0.21 μmol of the resulting compound of 59A were reacted in    the manner described in example 56B. After workup and purification    by preparative reversed-phase C₁₈ HPLC using a linear gradient of    35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant, 1.0 mg of the title    compound was obtained as a white solid. TLC: Rf=0.45, 3% MeOH in    CH₂Cl₂. HPLC: Rt=17.34 min; (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 60

Compound 60. To a solution of 70 mg of the resultant compound of Example51D in 10 mL of CH₂Cl₂ was added 3 mL of saturated aqueous sodiumbicarbonate solution, 50 mg of sodium bicarbonate, and 53 mg ofbenzofurazan-4-sulfonyl chloride. The mixture was stirred vigorously for4 h, then the resulting mixture was diluted with CH₂Cl₂, washed withsaturated brine, dried over magnesium sulfate and filtered. Afterconcentration of the mixture in vacuo, the residue was purified by thicklayer silica gel chromatography using 5% MeOH/CH₂Cl₂ as eluant to obtain80 mg of the title compound as a white solid. TLC: Rf=0.80, 5% MeOH inCH₂Cl₂. HPLC: Rt=14.96 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 61

Compound 61. To a solution of 35.5 mg (0.076 mmol) of the resultantcompound of Example 16 in 1 mL of dichloromethane was sequentially added27.6 μL (0.159 mmol) of diisopropylethyl amine and 12 μL (0.083 mmol) ofbenzyl chloroformate. After 1 hour, the mixture was concentrated invacuo. The residue was purified by preparative thin layer chromatographywith 50% ethyl acetate/dichloromethane as an eluent to yield 38.7 mg ofthe title compound as a white solid; TLC: Rf=0.63, 50% ethylacetate/dichlormethane; HPLC: Rt=15.45 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 62

-   A. Benzofurazan-4-sulfonic acid. To a solution of 252.0 mg (1.05    mmol) of o-nitroaniline-m-sulfonic acid sodium salt in 1 mL of water    was added 0.52 mL of 2.0 N HCl. After ½ h, 0.68 mL (1.05 mmol) of    tertrabutylammonium hydroxide (40% in water) was added. After 2    hours, the mixture was concentrated in vacuo. A solution of the    residue in 7 mL of acetic acid was treated with 488.5 mg (1.10 mmol)    of lead tertraacetate. After 24 hours, the precipitate was filtered    and washed with small amount of acetic acid. The solid was further    dried in vacuo to yield 267.9 mg of product. TLC: Rf=0.09, 10%    CH₃OH/CH₂Cl₂.-   B. Benzofurazan-4-sulfonyl chloride. To a solution of 137.0 mg    (0.522 mmol) of triphenylphosphine in 0.5 mL of dichloromethane was    slowly added 47 μL (0.594 mmol) of sulfuric chloride at 0° C. The    ice-water bath was removed and the crude resultant compound of    Example 62A in 0.5 mL of dichloromethane was added slowly. After 3    hours, the mixture was treated with 30 mL of 50% ether/hexane. The    supernatant was decanted into a dry flask and concentrated in vacuo.    The residue was purified by filtering through a plug of silica gel    with 25% ethyl acetate as an eluent to yield 23 mg of product. TLC:    Rf=0.6, 10% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with    structure.-   C. Compound 62. To a solution of 55.7 mg (0.166 mmol) of the    resultant compound of Example 39A in 1 mL of dichloromethane was    sequentially added 0.5 mL of saturated NaHCO₃, a small amount of    solid NaHCO₃ and the resultant compound of Example 62B. After 3    hours, the mixture was diluted with dichloromethane. The two layers    were separated and the aqueous layer was extracted once with    dichloromethane. The combined organic layer was washed with brine    then dried over MgSO₄, filtered and concentrated in vacuo. The    residue was purified by preparative HPLC to yield 5.3 mg of the    title compound as a white solid; TLC: Rf=0.40, 50% ethyl    acetate/dichloromethane; HPLC Rt=16.5 min; (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 63

-   A. A solution of 3.0 mg (0.0058 mmol) of the resultant title    compound of Example 62 in 2 mL ethyl acetate was treated with HCl    gas (moderate stream) for 3 minutes. The mixture was concentrated in    vacuo to yield the crude amine hydrochloride salt. TLC: Rf=0.20, 10%    CH₃OH/CH₂Cl₂.-   B. Compound 63. To a solution of the crude resultant compound of    Example 63A in 1 mL of dichloromethane was sequentially added 2.1 uL    (0.0121 mmol) of diisopropyl ethyl amine and 0.9 uL (0.0064 mmol) of    benzyl chloroformate. After 1 hour, the mixture was concentrated in    vacuo. The residue was purified by preparative thin layer    chromatography with 90% dichloromethane/methanol as an eluent to    yield 2.6 mg of the title compound as a white solid; TLC: Rf=0.34,    50% ethyl acetate/dichloromethane; HPLC, Rt=17.1 min; (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 64

-   A. 5-(Dimethylamino)thioxomethoxy)-benzofurazan. To a solution of    500 mg (3.67 mmol) of 5-hydroxybezofurazan in 10 mL of DMF was added    140 mg (4.59 mmol) of NaH in small portions. The resulting mixture    was stirred at room temperature until no more gas evolved. The flask    was then immersed in a cold water bath and 540 mg (4.41 mmol) of    dimethylthiocarbamoyl chloride (from Aldrich) was added. After 5    minutes, the water bath was removed the mixture was heated to 80° C.    for 1 hour. After being cooled to room temperature, the mixture was    poured into 20 mL of 0.5 N NaOH three times and water three times.    The solid was dried in vacuum to yield 580 mg of product that was    used in the next reaction without further purification; TLC:    Rf=0.20, 20% ethyl acetate/hexane; (¹H)-NMR (CDCl₃) consistent with    structure.-   B. 5-((Dimethylamino)carbonyl)thio)-benzofurazan. The crude product,    510 mg (2.28 mmol), from Example 64A was heated to 190° C. in a    sealed tube. After 5 hours, it was cooled to room temperature and    ethyl acetate was added. The solution was filtered through a plug of    a silica and concentrated in vacuo to yield 360 mg of product which    was again used in the next reaction without further purification.    TLC: Rf=0.20, 20% ethyl acetate/hexane.-   C. 5-Mercaptobenzofurazan. To a solution of 357.4 mg (1.60 mmol) of    the resultant compound of Example 64B in 2 mL of methanol was added    7 mL of 6 N NaOH. The mixture was heated to 90° C. for 2 hours. The    mixture was poured into 100 mL ice and acidified with concentrated    HCl. The slurry was filtered and rinsed three times with water. The    residue was dried in vacuo to yield 145.6 mg of product; TLC:    Rf=0.70, 20 ethyl acetate/hexane; (¹H)-NMR (CDCl₃) consistent with    structure.-   D. Benzofurazan-5-sulfonyl chloride. Chlorine gas was bubbled    through a solution of 39.9 mg (0.26 mmol) of the resultant compound    of Example 64C in a mixture of 1 mL of ethyl acetate and 0.5 mL of    water for 3 minutes. The mixture was then washed repeatedly with    brine until no more precipitate formed. The organic layer was dried    over MgSO₄, filtered and concentrated to yield 30 mg of the product    (52%). TLC: Rf=0.22, 20% ethyl acetate/hexane.-   E. Compound 64. A solution of the resultant compounds of Examples    52D and 39A (total yields) in a mixture of 1 mL of dichloromethane,    0.3 mL of saturated NaHCO₃ and a small amount of solid NaHCO₃ was    stirred at room temperature for 2 hours. The solution was diluted    with 30 mL of dichloromethane and the two layers were separated. The    aqueous layer was extracted once with dichloromethane chloride. The    combined organic layer was washed with brine, dried over MgSO₄ and    concentrated. The residue was purified by preparative thin layer    chromatography with 90% dichloromethane/ether as an eluent to yield    30 mg of the title product as a white solid; TLC: Rf=0.46, 10%    Et₂O/CH₂Cl₂, HPLC Rt=17.6 min; (¹H)-NMR (CDCl₃): δ8.45 (s), 1H;    7.96(d), 1H; 7.65 (d), 1H; 7.25(m), 5H; 4.65(d), 1H; 3.85(m), 1H;    3.78(m), 1H; 3.30(d), 2H; 3.10(m), 2H; 2.90(m), 2H: 1.90(m), 1H;    1.40(s), 9H; 0.90 (d), 6H.

EXAMPLE 65

Compound 65. A solution of 13.1 mg (0.025 mmol) of the resultantcompound of Example 64E in 1.5 mL of ethyl acetate was treated withgaseous HCl (moderate stream) at 0° C. for 3 minutes. The solvent wasremoved to yield a solid residue which was used in the next reactionwithout further purification; TLC: Rf=0.52, 10% CH₃OH/CH₂Cl₂. A solutionof this hydrochloride salt (entire yield) in 1 mL of dichloromethane wastreated sequentially with 9.2 μL (0.053 mmol) of diisopropyl ethyl amineand 4.0 μL (0.028 mmol) of benzyl chloroformate. After 3 hours, themixture was concentrated and purified by preparative thin layerchromatography with 90% dichloromethane/ether as an eluent to yield 11.7mg of the title compound as a white solid; TLC: Rf=0.65, 10%Et₂O/CH₂Cl₂; HPLC Rt=17.6 min; (¹H)-NMR (CDCl₃: δ 8.45(s), 1H; 7.96(d),1H; 7.65(d), 1H; 7.25(m), 10H; 5.00, (m), 2H; 4.85(d), 1H; 3.86(m), 2H;3.60(bs), 1H; 3.25(m), 12H; 3.05(d), 2H; 2.96(m), 1H: 2.98(m), 1H;1.88(m), 1H; 0.90(dd), 6H.

EXAMPLE 66

Compound 66. A solution of 100 mg (0.46 mmol) of the resultant compoundof Example 64D and 101 mg (0.286 mmol) of the resultant compound ofExample 48A in a mixture of 2 mL of dichloromethane, 0.5 mL of saturatedNaHCO₃ and small amount of solid NaHCO₃ was stirred at room temperaturefor 2 hours. The solution was diluted with 50 mL of dichloromethane andthe two layers were separated. The aqueous layer was extracted once withdichloromethane. The combined organic layer was washed with brine, driedover MgSO₄ and concentrated. The residue was purified by preparativethin layer chromatography with 20% ethyl acetate/hexane as an eluent toyield 82 mg of the title product as a slightly impure pale yellow solid.The material was further purified by preparative HPLC with a lineargradient solvent system of 35% to 80% of acetonitrile/water (0.1% TFA)over 80 min. Upon removal the solvents 50 mg of white solid wasobtained. TLC: Rf=0.46, 10% Et₂O/CH₂Cl₂; HPLC, Rt=17.6 min; (¹H)-NMR(CDCl₃): δ 8.45 (s), 1H; 7.96 (d), 1H; 7.65 (d), 1H; 7.25 (m), 5H; 5.15(m), 1H: 4.85 (d), 1H; 3.82 (m) 4H; 3.68 (d), 1H; 3.20(m), 2H, 3.05 (d),2H; 2.96 (m), 1H; 2.88 (m), 1H; 2.14(m), 1H; 1.92(m), 2H; 1.50(bs), 1H;0.90(dd), 6H.

EXAMPLE 67

Compound 67. Following the procedure described in Example 40B, asolution of the resultant compound of Example 40A in CH₂Cl₂ is treatedwith bis-((carboxamido)-amino)-acetic acid, diisopropylethylamine, HOBt,and EDC in a 1:1:1:1:1 molar ratio. the mixture is stirred for 16 h atambient temperature while protected from moisture, then diluted withadditional CH₂Cl₂ and washed sequentially with H₂O, saturated NaHCO₃solution and brine, then dried over MgSO₄ and concentrated in vacuo. Theresidue is purified by silica gel chromatography using an appropriateeluant to yield the title product.

EXAMPLE 68

Compound 68. This compound was prepared by the route described inExample 26, except that the reacting amine used was the resultingcompound of Example 39A (146 mg, 0.43 mmol) and the acylating agent was4-fluorophenyl sulphonyl chloride (27 mg, 0.14 mmol). Afterchromatographic purification on a silica gel column using 8%CH₃OH/CH₂Cl₂ as eluent, 92.8 mg of the title compound was obtained.HPLC: Rt=15.9 minutes. TLC: Rf=0.54, 8% MeOH/CH₂Cl₂; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 69

-   A. The resulting compound of Example 68 (72.1 mg, 0.167 mmol) was    dissolved in 90% aqueous TFA (3.3 mL), and the reaction mixture    stirred for 3 hours at room temperature, then was concentrated to    dryness. TLC: Rf=0.29, 8% MeOH/CH₂Cl₂.-   B. Compound 69. To a solution of the resulting compound of Example    69A (41.7 mg, 0.09 mmol) in CH₂Cl₂ (2 mL) was added    diisopropylethtylamine (47 μl, 0.27 mmol) and the resulting compound    of Example 48A (33 mg, 0.15 mmol), and the reaction proceeded for 14    hours at room temperature. The reaction mixture was then    concentrated, and the residue was chromatographed on a silica gel    column using 8% THF/CH₂Cl₂ was eluent, yielding the desired compound    which was further subjected to purification by preparative HPLC,    yielding 7.8 mg of a white solid. HPLC: Rt=13.5 minutes. TLC:    Rf=0.36, 8% THF/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 70

Compound 70. A solution of 30 mg of the resulting compound of Example 54and 17.6 mg of 3-acetamido-4-fluorobenzenesulfonyl chloride in 10 mL ofCH₂Cl₂ was reacted in the same manner as described for Example 14. Afterworkup and purification by preparative reversed-phase C₁₈ HPLC using alinear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 2.0 mgof the title compound was obtained. TLC: Rf=0.5, 10% CH₃OH in CH₂Cl₂.HPLC: Rt=13.74 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 71

Compound 71. A 30 mg portion of the resultant compound of Example 58 wasdeprotected with trifluoroacetic acid and the resulting compound reactedwith 9 μL of dimethysulfamoyl chloride in 10 mL of CH₂Cl₂ was reacted inthe manner described in Example 14. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 6.5 mg of the title compound wasobtained. TLC: Rf=0.2, 3% MeOH in CH₂Cl₂. HPLC: Rt=15.96 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 72

Compound 72. A solution of the resulting compound from thetrifluoroacetic acid deprotection of Example 69A (31 mg, 0.07 mmol) inCH₂Cl₂ (2 mL) was added diisopropylethylamine (47 μl, 0.27 mmol) anddimethylsulfamoyl chloride (22 μl, 0.20 mmol), and the reactionproceeded for 16 hours at room temperature. The reaction mixture wasthen concentrated, and the residue was chromatographed on a thick layersilica gel plate (1.0 mm) using 5% THF/CH₂Cl₂ as eluent, yielding thedesired compound which was further subjected to purification bypreparative HPLC to yield 7.8 mg of a white solid. HPLC: Rt=14.8minutes. TLC: Rf=0.44, 5% THF/CH₂Cl₂.

EXAMPLE 73

Compound 73. A 43 mg portion of the resultant compound of Example 54 wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 12 h. Themixture was concentrated in vacuo and the residue taken up in 5 mL ofCH₂Cl₂. To this solution, 3 mL saturated aqueous sodium bicarbonate and25 mg of 2,5-dimethoxybenzenesulfonyl chloride was added, and themixture was stirred for 12 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bythick layer silica gel chromatography using 3% MeOH/CH₂Cl₂ as eluantfollowed by preparative reversed-phase C₁₈ HPLC using a linear gradientof 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain 5.5 mg of thetitle compound. TLC: Rf=0.20, 3% MeOH/CH₂Cl₂. HPLC: Rt=15.15 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 74

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=cyclopropylmethyl, A′=H).    To a solution of compound XX (A=tert-butoxycarbonyl) (0.8 g, 2.67    mmol) in ethanol (30 mL) was added a solution of KOH (0.18 g, 3.2    mmol) in ethanol (20 mL) and the mixture stirred for 45 min at room    temperature. In a separate flask, a solution of    cyclopropylmethyl-amine hydrochloride (1.44 g, 13.3 mmol) in ethanol    (20 mL) was added KOH (0.75 g, 13.3 mmol). The mixture was stirred    30 min at room temperature. The solutions were combined and heated    at 85° C. for 3 h. The solution was concentrated under reduced    pressure and the residue slurried in diethyl ether and filtered. The    ethereal layer was concentrated to give 0.32 g of a white solid;    (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound 74. To a solution of the resulting compound of Example    74A (0.1 g, 0.30 mmol) in CH₂Cl₂ (20 mL) was added a saturated    solution of sodium bicarbonate, followed by addition of solid sodium    bicarbonate (30 mg, 0.36 mmol), then 4-fluorobenzenesulfonyl    chloride (0.07 g, 0.36 mmol). The mixture was allowed to stir at    room temperature for 4 h. The organics were extracted into 250 mL    CH₂Cl₂, dried over anhydrous MgSO₄, concentrated under reduced    pressure then purified via medium pressure liquid chromatography    using a gradient system of CH₂Cl₂ followed by 0.5:99.5    methanol/CH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂. The title compound    was obtained as 35 mg of a colorless foam. HPLC: Rt=16.8 min. TLC:    Rf=0.32, 3:97 methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 75

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=isopropyl, A′=H). To a    solution of Compound XX (A=tert-butoxycarbonyl) (1.67 mmol) in    ethanol (10 mL) was treated with isopropylamine (10 mL). The    solution was heated to 85° C. for 72 h. The solution was filtered    then concentrated under reduced pressure to give 0.56 g of the title    compound which was used without subsequent purification. (¹H)-NMR    (CDCl₃) consistent with structure.-   B. Compound 75. To a solution of the resultant compound of Example    75A (0.2 g, 0.65 mmol) in CH₂Cl₂ (10 mL) was added a saturated    solution of sodium bicarbonate (3 mL), followed by addition of solid    sodium bicarbonate (0.11 g, 1.31 mmol), then p-fluorobenzenesulfonyl    chloride (0.25 g, 1.28 mmol). The mixture was stirred overnight at    ambient temperature. The organics were extracted into 100 mL CH₂Cl₂,    dried over anhydrous MgSO₄, concentrated under reduced pressure then    purified via medium pressure silica gel chromatography using a    gradient system of CH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂. The    title compound was obtained as a colorless foam 200 mg. TLC:    Rf=0.22, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.48 min; (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 76

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=morpholinyl, A′=H). To a    solution of compound XX (A=Boc) in ethanol is added 3 molar    equivalents of N-amino morpholine. The mixture is heated under    reflux for 12 h, cooled, and the mixture concentrated in vacuo. The    residue is purified by preparative reversed-phase chromatography    using a linear gradient of 5% to 100% acetonitrile/H₂O as eluant to    yield the title compound.-   B. Compound 76. Following the procedure described in Example 81, a    solution of the resultant compound of Example 76A in CH₂Cl₂ is    reacted with 4-fluorobenzenesulfonyl chloride in the presence of    water and NaHCO₃. Following dilution with additional CH₂Cl₂ and    aqueous workup, the resultant product is dried over MgSO₄, filtered,    and concentrated in vacuo. The residue is then purified by silica    gel chromatography using an appropriate solvent system to yield the    title product.

EXAMPLE 77

-   A. Compound XXI (A=tert-butoxycarbonyl,    D′=4-(N,N-dimethylamino)-benzyl, A′=H). To a solution of compound XX    (A=Boc) in ethanol is added 3 molar equivalents of    4-aminomethyl-(N,N-dimethyl)-aniline. The mixture is heated under    reflux for 12 h, cooled, and the mixture concentrated in vacuo. The    residue is purified by silica gel chromatography using an    appropriate solvent system as eluant to yield the title product.-   B. Compound 77. Following the procedure described in Example 81, a    solution of the resultant compound of Example 77A in CH₂Cl₂ is    reacted with 4-fluorobenzenesulfonyl chloride in the presence of    water and NaHCO₃. Following dilution with additional CH₂Cl₂ and    aqueous workup, the resultant product is dried over MgSO₄, filtered,    and concentrated in vacuo. The residue is then purified by silica    gel chromatography using an appropriate solvent system to yield the    title product.

EXAMPLE 78

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=cyclopentyl, A′=H). To a    solution of compound XX (A=Boc) in ethanol is added 10 molar    equivalents of cyclopentylamine. The mixture is heated under reflux    for 12 h, cooled, and the mixture concentrated in vacuo. The residue    is used without subsequent purification.-   B. Compound 78. Following the procedure described in Example 81, a    solution of the resultant compound of Example 78A in CH₂Cl₂ is    reacted with 4-fluorobenzenesulfonyl chloride in the presence of    water and NaHCO₃. Following dilution with additional CH₂Cl₂ and    aqueous workup, the resultant product is dried over MgSO₄, filtered,    and concentrated in vacuo. The residue is then purified by silica    gel chromatography using an appropriate solvent system to yield the    title product.

EXAMPLE 79

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=2-(4-pyridyl)ethyl,    A′=H). To a solution of compound XX (A=Boc) in ethanol is added 3    molar equivalents of 4-aminoethylpyridine. The mixture is heated    under reflux for 12 h, cooled, and the mixture concentrated in    vacuo. The residue is purified by preparative reversed-phase    chromatography using a linear gradient of 5% to 100%    acetonitrile/H₂O as eluant to yield the title product.-   B. Compound 79. Following the procedure described in Example 81, a    solution of the resultant compound of Example 79A in CH₂Cl₂ is    reacted with 4-fluorobenzenesulfonyl chloride in the presence of    water and NaHCO₃. Following dilution with additional CH₂Cl₂ and    aqueous workup, the resultant product is dried over MgSO₄, filtered,    and concentrated in vacuo. The residue is then purified by silica    gel chromatography using an appropriate solvent system to yield the    title product.

EXAMPLE 80

-   A. 4-Cyanotetrahydro-4H-pyran. Following essentially the procedure    of Yoneda, R. “Cyanophosphate: An Efficient intermediate for    Conversion of Carbonyl compounds to Nitriles,” Tetrahedron Lett.,    30, 3681 (1989), a solution of tetrahydro-4H-pyran-one (9.9 g, 97.8    mmol) in dry THF (50 mL) is reacted with lithium cyanide (9.7 g, 294    mmol) and diethylcyanophosphonate (24 g, 146 mmol). The mixture is    stirred for 24 h at ambient temperature. The reaction is quenched by    the addition of 100 mL H₂O. The product is extracted into 1.5 L of    diethyl ether, dried over anhydrous MgSO₄ then concentrated under    reduced pressure. The residue is dissolved in dry THF (30 mL) and    tert-butyl alcohol (7.25 g, 97.8 mmol). This solution is added    slowly to 75 mL of a 1 M solution of SmI₂. The mixture is stirred    for 15 h at ambient temperature. The reaction is quenched by    addition of 100 mL of saturated aqueous NH₄Cl. The resulting mixture    is extracted with diethyl ether and the organic layers dried over    anhydrous MgSO₄ and concentrated under reduced pressure.    Purification by silica gel chromatography gives the title compound.-   B. 4-(aminomethyl)tetrahydro-4H-pyran To a solution of the compound    of the Example 80A (10 g, 89.9 mmol) in absolute ethanol (200 mL) is    added Raney Nickel (2.0 g, 50% slurry in water). The mixture is    stirred for 24 hours at ambient temperature under 40 psig of    hydrogen. The solution is filtered through celite and the solution    concentrated under reduced pressure. The residue is taken up in    ether (2L) washed with brine, dried in anh. MgSO₄, then concentrated    under reduced pressure to give the title commpound.-   C.    (1S,2R)-N-(1-Benzyl-3-(N-(4-(aminomethyl)tetrahydro-4H-pyran))-2-hydroxypropyl)-tert    butoxycarbonylamine. To a solution of the compound of Example 80B (5    g, 48.5 mmol) in absolute ethanol (20 mL) is added the compound XX    (A=Boc)(2.55 g, 9.7 mmol). The mixture is stirred for 24 hours at    ambient temperature. The solution is concentrated under reduced    pressure and the crude product is puffed via column chromatography    to give the title compound.-   D. Compound XXII (A=Boc, D′=(4-tetrahydro-4H-pyranyl)methyl, A′=H).    To a solution of compound XX (A=Boc) in ethanol is added 3 molar    equivalents of the resulting compound of Example 80° C. The mixture    is heated under reflux for 12 h, cooled, and the mixture    concentrated in vacuo. The residue is purified by preparative    reversed-phase chromotography using a linear gradient of 5% to 100%    acetonitrile/H₂O as eluant to yield the title compound.

To a solution of compound XX(A=Boc) in ethanol is added 3 molarequivalents of N-amino morpholine. The mixture is heated under refluxfor 12 h, cooled, and the mixture concentrated in vacuo. The residue ispurified by preparative reversed-phase chromatography using a lineargradient of 5% to 100% acetonitrile/H₂O as eluant to yield the titlecompound.

-   E. Compound 80. Following the procedure described in Example 81, a    solution of the resultant compound of Example 80D in CH₂Cl₂ is    reacted with 4-fluorobenzenesulfonyl chloride in the presence of    water and NaHCO₃. Following dilution with additional CH₂Cl₂ and    aqueous workup, the resultant product is dried over MgSO₄, filtered,    and concentrated in vacuo. The residue is then purified by silica    gel chromatography to yield the title product.

EXAMPLE 81

-   A. Compound XXII (P=tert-butoxycarbonyl, D′=isobutyl,    E=3,4-dichlorophenyl). A solution of 316 mg of the resultant    compound of Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was    treated sequentially, at ambient temperature under an atmosphere of    nitrogen, with 276 mg of 3,4-dichorobenzenesulfonyl chloride and 95    mg of sodium bicarbonate. The mixture was stirred for 14 h, diluted    with CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ as    eluent to yield 490 mg of product. TLC: Rf=0.26, 5% diethyl ether in    CH₂Cl₂. HPLC: Rt=18.92 min. (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound XXII (P=H, D′=isobutyl, E=3,4-dichlorophenyl,    hydrochloride salt). A solution of 467 mg of the resultant compound    of Example 81A in ethyl acetate was treated at −20° C. with HCl gas.    The HCl was bubbled through the mixture for 20 min over which time    the temperature was allowed to warm to 20° C. Nitrogen was then    bubbled through the mixture for 15 min and solvent removed in vacuo    to yield 412 mg of product as a white solid which was used without    subsequent purification.-   C. Compound 81. A solution of 91 mg of the resultant compound of    Example 81B in CH₂Cl₂ was treated sequentially, at ambient    temperature under an atmosphere of nitrogen, with 25 mg of allyl    chloroformate and 52 mg N,N-diisopropylethylamine. The mixture was    stirred for 4 h and then concentrated in vacuo. The residue was    taken up in ethyl acetate and washed with 0.5 N HCl and saturated    NaCl then dried over MgSO₄, filtered, and concentrated in vacuo to    yield 89 mg of the title product as a white solid. TLC: Rf=0.53, 5%    diethyl ether in CH₂Cl₂. HPLC: Rt=17.95 min. (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 82

-   A. (3-Pyridyl)-methyl-4-nitrophenyl-carbonate. To a solution of 3.65    g of bis-(nitrophenyl) carbonate in 25 mL of CH₂Cl₂ at 0° C. was    added sequentially 0.97 mL of 3-pyridyl carbinol and 1.3 mL of    4-methyl morphine. After stirring at room temperature for 24 hours,    the resultant mixture was diluted with 100 mL of CH₂Cl₂, washed with    saturated sodium bicarbonate, water and brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by filtration through a plug of silica gel, using 0-40%    EtOAc/CH₂Cl₂ as eluent to provide 1.68 g of the title product. TLC:    Rf=0.19, 50% EtOAc/hexane.-   B. Compound XXII (P=tert-butoxycarbonyl, D′=isobutyl,    E=3,4-benzofurazan). To a solution of 498.6 mg of the resultant    compound of Example 39A in 10 mL of CH₂Cl₂ was added sequentially, 2    mL of saturated sodium bicarbonate, a small amount of solid sodium    bicarbonate and 518.4 mg of the resultant compound of Example 64D.    After stirring at room temperature for 3 hours, the resultant    mixture was diluted with 60 mL of CH₂C₂, washed with saturated    sodium bicarbonate and brine, dried over magnesium sulfate, filtered    and concentrated in vacuo. The residue was purified by silica gel    chromatography using 5% diethyl ether/hexane as eluent to yield 300    mg of white solid. TLC: Rf=0.80, 50% EtOAc/hexane.-   C. Compound XXII (P=H, D′=isobutyl; E=3,4-benzofurazan,    hydrochloride salt.). A solution of 60.3 mg of the resultant    compound of Example 82B in 3 mL of EtOAc at −20° C. was treated with    anhydrous HCl gas for 5 min. The ice bath was removed and after an    additional 10 min. The reaction mixture was sparged with nitrogen    then concentrated in vacuum and the resulting white solid used    without subsequent purification for subsequent reaction.-   D. Compound 82. To a solution of the resultant compound of Example    82C (entire yield) in 2 mL of CH₂Cl₂ was added sequentially, 45 μL    of diisopropylethylamine and 35.1 mg of the resultant compound of    Example 82A. The mixture was stirred for 24 hours and then    concentrated in vacuo. The residue was purified by preparative thin    layer chromatography using 60% ether/CH₂Cl₂ as eluent followed by    preparative reversed-phase C₁₈ HPLC using a linear gradient of 40%    to 100% CH₃CN/H₂O with 0.1% TFA as eluant. The resultant TFA salt of    the title compound was washed with saturated sodium bicarbonate to    yield 6.5 mg of the title compound. TLC: Rf=0.15, 20% EtOAc/CH₂Cl₂.    HPLC: Rt=13.52 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 83

-   A. Compound XXII (A=tert-butoxycaronyl, D′=isobutyl,    E=4-acetamido-3-chlorophenyl). A solution of 339 mg of the resultant    compound of Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was    treated sequentially, at ambient temperature under an atmosphere of    nitrogen, with 324 mg of 4-acetamido-3-chlorobenzenesulfonyl    chloride and 102 mg of sodium bicarbonate. The mixture was stirred    for 14 h, diluted with CH₂Cl₂, washed with saturated NaCl then dried    over MgSO₄, filtered, and concentrated in vacuo. The residue was    purified by low pressure silica gel chromatography using 20% diethyl    ether in CH₂Cl₂ as eluent to yield 498 mg of product. TLC: Rf=0.27    (20% diethyl ether in CH₂Cl₂). HPLC: Rt=16.20 min. (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (A=H, D′=isobutyl, E=4-acetamido-3-chlorophenyl,    hydrochloride salt). A solution of 474 mg of the resultant compound    of Example 83A in ethyl acetate was treated at −20° C. with HCl gas.    The HCl was bubbled through the mixture for 20 min over which time    the temperature was allowed to warm to 20° C. Nitrogen was then    bubbled through the mixture for 15 min and the solvent was removed    in vacuo to yield 421 mg of product as a white solid which was used    without subsequent purification.-   C. Compound 83. A solution of 92 mg of the resultant compound of    Example 83B in CH₂Cl₂ was treated sequentially, at ambient    temperature under an atmosphere of nitrogen, with 24 mg of allyl    chloroformate and 52 mg N,N-diisopropylethylamine. The mixture was    stirred for 4 h and then concentrated in vacuo. The residue was    taken up in ethyl acetate and washed with 0.5 N HCl and saturated    NaCl, then dried over MgSO₄, filtered, and concentrated in vacuo to    yield 106 mg of the title product as a white solid. TLC: Rf=0.38    (20% diethyl ether in CH₂Cl₂). HPLC: Rt=15.28 min. (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 84

Compound XXII (P=tert-butoxycarbonyl, D′=isobutyl,E=3,4-dichlorophenyl). To a solution of the resultant compound ofExample 51D (220 mg, 0.61 mmol) in CH₂Cl₂ (10 mL) was added the3,4-dichlorobenzenesulfonyl chloride (300 mg, 1.22 mmol) followed by theaddition of a saturated solution of sodium bicarbonate (3 mL) followedby addition of 0.1 g of solid sodium bicarbonate. The mixture wasstirred at ambient temperature overnight. The solution was diluted with100 mL CH₂Cl₂, the organics separated, dried over anhydrous, MgSO₄, andthe organics concentrated under reduced pressure to obtain 0.17 g ofcrude product. The crude product was purified via medium pressure liquidchromatography using CH₂Cl₂ followed by 0.5:99.5 methanol/CH₂Cl₂followed by a 1:99 methanol/CH₂Cl₂ solution as the solvent system togive 103 mg of the title compound as a white solid. TLC: Rf=0.56 (3:97methanol/CH₂Cl₂), HPLC: Rt=19.78 min, (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 85

-   A. (3-Tetrahydrofuryl)-methyl-4-nitrophenyl-carbonate. To a solution    of 1.21 g of p-nitrophenyl chloroformate in 20 mL of CH₂Cl₂ 0° C.    was added sequentially, 0.51 g of tetrahydro-3-furanmethanol and    0.66 mL of 4-methyl morpholine. After stirring at room temperature    for 2 hours. The mixture was stirred for 2 hours and concentrated in    vacuo. The residue was purified by filtering through a plug of    silica gel, using 0-50% EtOAc/CH₂Cl₂ as eluent to provide 1.17 g of    the title product as a pale yellow solid. TLC: Rf=0.20, 50%    EtOAc/hexane.-   B. Compound 85. To a solution of 70 mg of the resultant compound of    Example 81B in 1 mL of THF was added sequentially, 56 μL of    diisopropylethylamine and a solution of 46.6 mg of the resultant    compound of Example 85A in 1 mL of THF. The mixture was stirred for    24 hours and then concentrated in vacuo. The residue was diluted    with 60 mL of CH₂Cl₂, washed with 5% sodium bicarbonate and brine,    dried over magnesium sulfate, filtered and concentrated in vacuo to    yield 120 mg of crude product. The residue was purified by    preparative thin layer chromatography using 20% EtOAc/CH₂Cl₂ as    eluent to yield 82 mg of the title compound. TLC: Rf=0.4, 20%    EtOAc/CH₂Cl₂. HPLC: Rt=17.08 min. (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 86

Compound 86. A solution of 42 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 41 mg of the product of Example 52A and 46mg N,N-diisopropylethylamine. The mixture was stirred 14 h, diluted withCH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl, then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using ethyl acetate aseluent to yield 43 mg of product. TLC: Rf=0.44 (20% ethyl acetate).HPLC: Rt=13.14 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 87

-   A. Compound XXII (P=H, D′=isobutyl, E=4-acetamido, 3-fluoro). A    solution of 25 mg of the resultant compound of Example 54 in EtOAc    (10 mL) at 0° C. was treated with anhydrous hydrogen chloride gas    for 10 min., and allowed to stand for 12 h while warming to ambient    temperature. The resultant mixture was then concentrated in vacuo to    yield compound as a white solid which was used without subsequent    purification for ensuing reaction.-   B. Compound 87. A 0.045 mmol portion of the resultant compound of    Example 87A was taken up in 5 mL of CH₂Cl₂. To this solution, 40 μL    of diisopropylethylamine and 6 μL of allyl chloroformate were added    at 0° C. and the mixture was stirred for 12 h, while warming slowly    to ambient temperature. The resulting mixture was diluted with    CH₂Cl₂, washed with saturated brine, dried over magnesium sulfate    and filtered. After concentrated in vacuo, the residue was purified    by preparative reversed-phase C₁₈ HPLC using a linear gradient of    35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain 11.6 mg of    the title compound. TLC: Rf=0.20, 5% MeOH/CH₂Cl₂. HPLC: Rt=14.6 min;    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 88

Compound 88. A 0.033 mmol portion of the resultant compound of Example87A was taken up in 5 mL of CH₂Cl₂. To this solution, 26 μL oftriethylamine and 12 mg of the resultant compound of Example 48A wereadded and stirred for 12 h. The resulting mixture was diluted withCH₂Cl₂, washed with saturated sodium bicarbonate solution and saturatedbrine, dried over magnesium sulfate and filtered. After concentration ofthe mixture in vacuo, the residue was purified by thick layer silica gelchromatography using 5% MeOH/CH₂Cl₂ as eluant followed by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluant to obtain 7.5 mg of the title compound. TLC:Rf=0.30, 5% MeOH/CH₂Cl₂. HPLC: Rt=13.38 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 89

Compound 89. A solution of 28 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of n-propyl chloroformate and 17 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 31 mg of the title productas a white solid. TLC: Rf=0.35 (5% diethyl ether in CH₂Cl₂). HPLC:Rt=18.12 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 90

Compound 90. A solution of 28 mg of the resultant compound of Example83B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 7 mg of n-propyl chloroformate and 15 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 30 mg of the title productas a white solid. TLC: Rf=0.47 (20% diethyl ether in CH₂Cl₂). HPLC:Rt=15.41 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 91

-   A. 3-Acetamidobenzene sulfonic acid. A solution of 1.48 g of    3-aminobenzene sulfonic acid in 1:1 tetrahydrofuran/water was    treated at 0° C. with 1.43 g of sodium bicarbonate. After 5 min,    1.30 g of acetic anhydride was added dropwise and the reaction    allowed to warm to ambient temperature under an atmosphere of    nitrogen over 14 h. The reaction mixture was passed through a column    of Amberlyst 15 ion exchange resin, eluted with water, and    concentrated in vacuo to yield an oil which upon treatment with    benzene and azeotropic removal of water in vacuo yielded 1.8 g of    the title product as a white crystalline solid. (¹H)-NMR (CDCl₃)    consistent with structure.-   B. 3-Acetamidobenzene sulfonic acid, sodium salt. The resultant    compound of Example 91A in water was treated at 0° C. with 8.5 mL of    1 N sodium hydroxide. The mixture was stirred for 3 h and    concentrated in vacuo to yield an oil which upon treatment with    benzene and azeotropic removal of water in vacuo yielded the title    product as a tan solid which was used directly in the next reaction.-   C. 3-Acetamidobenzenesulfonyl chloride. The resultant compound of    Example 91B in CH₂Cl₂ was treated at 0° C. with 4.5 g of phosphorous    pentachloride under an atmosphere of nitrogen. The mixture was    stirred 14 h, extracted with CH₂Cl₂, and concentrated in vacuo to    yield 1.7 g of the title product as a brown oil. TLC: Rf=0.21 (1:1    toluene/diethyl ether). (¹H)-NMR (CDCl₃) consistent with structure.-   D. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,    E=3-acetamidophenyl). A solution of 280 mg of the resultant compound    of Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 252 mg of the resultant compound of Example 91C and    105 mg of sodium bicarbonate. The mixture was stirred for 60 h,    diluted with CH₂Cl₂, washed with saturated NaCl then dried over    MgSO₄, filtered and concentrated in vacuo. The residue was purifed    by low pressure silica gel chromatography using 20% diethyl ether in    CH₂Cl₂ as eluent to yield 156 mg of the title product. TLC: Rf=0.14    (20% diethyl ether in CH₂Cl₂was). HPLC: Rt=15.39 min. (¹H)-NMR    (CDCl₃) consistent with structure.-   E. Compound XXII (A=H, D′=isobutyl, E=3-acetamidophenyl,    hydrochloride salt). A solution of 123 mg of the resultant compound    of Example 91D in ethyl acetate was treated at −20° C. with HCl gas.    The HCl was bubbled through the mixture for 20 min, over which time    the temperature was allowed to warm to 20° C. Nitrogen was then    bubbled through the mixture for 15 min and solvent removed in vacuo    to yield 118 mg of the title product as a white solid which was used    directly in subsequent reactions.-   F. Compound 91. A solution of 49 mg of the resultant compound of    Example 91E in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 48 mg of the resultant    compound of Example 48A and 54 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 14 h, diluted with CH₂Cl₂,    washed with saturated NaHCO₃ and saturated NaCl, then dried over    MgSO₄, filtered and concentrated in vacuo. The residue was subjected    to preparative thin layer silica gel chromatography using 5% CH₃OH    in CH₂Cl₂ to yield 42 mg of product. TLC: Rf=0.32 (5% CH₃OH in    CH₂Cl₂). HPLC: Rt=13.27 min. (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 92

Compound 92. To a solution of 63.5 mg of the resultant compound ofExample 17B, diastereomer B in 1 mL of THF was added sequentially, 52 μLof diisopropylethylamine and a solution of 43.3 mg of the resultantcompound of Example 85A in 1 mL of THF. The mixture was stirred for 24hours and then concentrated in vacuo. The residue was diluted with 60 mLof CH₂Cl₂, washed with 5% sodium bicarbonate and brine, dried overmagnesium sulfate, filtered and concentrated in vacuo to yield 70.7 mgof crude product. The residue was purified by preparative reversed-phaseC₁₈ HPLC using a linear gradient of 30% to 100% CH₃CN/H₂O with 0.1% TFAas eluant to obtain 43.9 mg of the title compound. TLC: Rf=0.29, 100%EtOAc. HPLC: Rt=13.24 min; (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 93

-   A. N-hydroxysuccinimidyl-(R)-3-hydroxytetrahydrofuryl carbonate. The    title compound was prepared as described in Example 48A starting    with 81 mg of (R)-3-hydroxytetrahydrofuran to yield 56 mg of the    title product as a white solid. (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound 93. To a solution of 43 mg of the resultant compound of    Example 35A in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, 27 mg of the resultant compound of Example    93A and 39 mg N,N-diisopropylethylamine. The mixture was stirred for    14 h, diluted with CH₂Cl₂, washed with saturated NaHCO₃ and    saturated NaCl, then dried over MgSO₄, filtered, and concentrated in    vacuo. The residue was purified by preparative thin layer silica gel    chromatography using 2% CH₃OH in CH₂Cl₂ as eluent to yield 45 mg of    the title product as a white solid. TLC: Rf=0.52 (5% CH₃OH CH₂Cl₂).    HPLC: Rt=14.94 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 94

Compound 94. A solution of 47 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 28 mg of the product of Example 93A and 39mg N,N-diisopropylethylamine. The mixture was stirred for 14 h, dilutedwith CH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl, then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by preparative thin layer silica gel chromatography using 5%methanol in CH₂Cl₂ as eluent to yield 40 mg of the title product as awhite solid. TLC: Rf=0.38 (ethyl acetate). HPLC: Rt=13.09 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 95

Compound 95. To a solution of 72.0 mg (0.189 mmol) of the resultantcompound of Example 51D in CH₂Cl₂ (4 mL) was added aqueous sodiumbicarbonate (1 mL), solid sodium bicarbonate 19.1 mg (0.227 mmol), and2,3-dichlorothiophenesulfonyl chloride 57.1 mg, (0.227 mmol). After 14h, the resulting mixture was diluted with EtOAc, washed with saturatedbrine, dried over magnesium sulfate, filtered and concentrated in vacuo.The residue was purified by low pressure silica gel columnchromatography using 5 to 12% EtOAc/CH₂Cl₂ eluent to provide 49.1 mg ofthe title product. TLC: Rf=0.62 25% EtOAc/CH₂Cl₂, HPLC: Rt=17.3 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 96

-   A. (4-Acetamido)-phenylmethyl-4-nitrophenyl-carbonate. To a solution    of 242.8 mg of p-nitrophenyl chloroformate in 5 mL of acetonitrile    at 0° C. was added sequentially, 165.2 mg of 4-acetamidobenzyl    alcohol and 0.13 mL of 4-methyl morpholine. The mixture was stirred    for 24 hours and concentrated in vacuo. The residue was taken up in    CH₂Cl₂ and washed with 5% sodium bicarbonate and brine, dried over    magnesium sulfate, filtered and concentrated in vacuo to yield 320    mg of the title compound. TLC: Rf=0.23, 50% EtOAc/hexane.-   B. Compound 96. To solution of the resultant compound of Example 40A    in 1 mL of THF was added sequentially, 56 μL of    diisopropylethylamine and 63 mg of the resultant compound of Example    96A. The mixture was stirred for 24 hours and then concentrated in    vacuo. The residue was purified by preparative thin layer    chromatography using 10% methanol/CH₂Cl₂ as eluent followed by    preparative reversed-phase C₁₈ HPLC using a linear gradient of 30%    to 100% CH₃CN/H₂O with 0.1% TFA as eluant to yield 50.2 mg of the    title compound. TLC: Rf=0.43, 10% methanol/CH₂Cl₂. HPLC: Rt=13.54    min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 97

Compound 97. To solution of 60 mg of the resultant compound of Example35A in 1 mL of THF was added sequentially, 54 μL ofdiisopropylethylamine and a solution of 48.9 mg of the resultantcompound of Example 85A in 1 mL THF. The mixture was stirred for 24hours and then concentrated in vacuo. The residue was diluted with 60 mLof CH₂Cl₂, washed with 5% sodium bicarbonate and brine, dried overmagnesium sulfate, filtered and concentrated in vacuo. The residue waspurified by preparative thin layer chromatography using 20% EtOAc/CH₂Cl₂as eluent to yield 46.9 mg of the title compound. TLC: Rf=0.31, 20%EtOAc/CH₂Cl₂. HPLC: Rt=15.18 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 98

Compound 98. To a solution of 61.0 mg of the resultant compound ofExample 35A in 1 mL of THF was added sequentially, 49 μL ofdiisopropylethylamine and a solution of 44 mg of the resultant compoundof Example 82A in 1 mL THF. The mixture was stirred for 24 hours andthen concentrated in vacuo. The residue was purified by preparative thinlayer chromatography using 5% methanol/CH₂Cl₂ as eluent to yield 61.0 mgof a white solid. TLC: Rf=0.19, 5 methanol/CH₂Cl₂. HPLC: Rt=13.28 min;13.28 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 99

Compound 99. A solution of 75 mg of the resultant compound of Example51D and 45 mg of 4-chlorobenzenesulfonyl chloride were reacted in themanner described in Example 60. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 24.6 mg of the title compoundwas obtained. TLC: Rf=0.3, 4% MeOH/CH₂Cl₂. HPLC: Rt=15.87 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 100

Compound 100. A solution of 40 mg of the resultant compound of Example51D and 45 mg of 4-methoxybenzenesulfonyl chloride were reacted in themanner described in Example 60. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 21.4 mg of the title compoundwas obtained as a white solid. TLC: Rf=0.2, 4% MeOH/CH₂Cl₂. HPLC:Rt=14.85 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 101

Compound 101. This compound was prepared from the resultant compound ofExample 128 by treatment with hydrogen chloride gas and subsequentreaction with the resultant compound of Example 48A in the mannerdescribed in Example 132. After workup and purification by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluant on a portion of the crude mixture, 4.2 mg of thetitle compound was obtained as a white solid. TLC: Rf=0.2, 4%MeOH/CH₂Cl₂. HPLC: Rt=11.53 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 102

Compound 102. A solution of 36 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of methyl chloroformate and 22 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 30% diethyl ether in CH₂Cl₂ aseluent to provide 27 mg of the title product as a white solid. TLC:Rf=0.10 (30% diethyl ether in CH₂Cl₂). HPLC: Rt=13.49 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 103

Compound 103. A solution of 29 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 6 mg of methyl chloroformate and 17 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 29 mg of the title product as a white solid. TLC:Rf=0.24 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=17.07 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 104

Compound 104. A solution of 31 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of methyl chloroformate and 21 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 24 mg of the title product as a white solid. TLC:Rf=0.23 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=15.41 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 105

-   A. N-hydroxysuccinimidyl methallyl carbonate. To a solution of 2.9    mL of 1.93 M phosgene in toluene at −10° C. was added 857 mg of    methallyl alcohol. The mixture was stirred for 2 h at −10° C. to    produce a 1.9 M solution of the title compound which was used    directly in subsequent reactions.-   B. Compound 105. A solution of 39 mg of the resultant compound of    Example 40A in CH₂Cl₂ was treated sequentially, at ambient    temperature under an atmosphere of nitrogen, with 0.05 mL of the    resultant compound of Example 105A and 24 mg    N,N-diisopropylethylamine. The mixture was stirred for 3 h and then    concentrated in vacuo. The residue was taken up in ethyl acetate and    washed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by    preparative thin layer silica gel chromatography using ethyl acetate    as eluent to yield 18 mg of the title product as a white solid. TLC:    Rf=0.67 (ethyl acetate). HPLC: Rt=14.97 min. (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 106

Compound 106. A solution of 31 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 0.04 mL of the resultant compound ofExample 105A and 18 mg N,N-diisopropylethylamine. The mixture wasstirred for 3 h and then concentrated in vacuo. The residue was taken upin ethyl acetate and washed with 0.5 N HCl and saturated NaCl then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 5% diethylether/CH₂Cl₂ as eluent to provide 19 mg of the title product as a whitesolid. TLC: Rf=0.34 (5% diethyl ether/CH₂Cl₂). HPLC: Rt=18.24 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 107

Compound 107. A solution of 28 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 0.05 mL of the resultant compound ofExample 105A and 19 mg N,N-diisopropylethylamine. The mixture wasstirred for 3 h and then concentrated in vacuo. The residue was taken upin ethyl acetate and washed with 0.5 N HCl and saturated NaCl then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 5% diethylether in CH₂Cl₂ as eluent to provide 18 mg of the title product as awhite solid. TLC: Rf=0.25 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=16.68min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 108

Compound 108. To a solution of 62.5 mg of 124B in 1 mL of THF was addedsequentially 56 μL of diisopropylethylamine and a solution of 49.6 mg ofthe resultant compound of Example 82A in 1 mL THF. The mixture wasstirred for 24 hours and then concentrated in vacuo. The residue waspurified by preparative thin layer chromatography using 50% EtOAc/CH₂Cl₂as eluent followed by preparative reversed-phase C18 HPLC using a lineargradient of 30% to 100% CH₃CN/H₂O with 0.1% TFA as eluant on a portionof the crude mixture, 4.2 mg of the title compound was obtained as awhite solid. TLC: Rf=0.16, 10% methanol/CH₂Cl₂. HPLC: Rt=13.67 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 109

-   A. (S)-4-Methoxycarbonyl-oxazlidin-2-one. To a solution of 4.88 g of    serine methyl ester hydrochloride in 25 mL of water was added 6.94 g    of potassium carbonate. The mixture was cooled to 0° C. and 19.5 mL    of phosgene was added dropwise. After stirring at 0° C. for 3 hours,    water was removed to yield a white solid with was washed with    copious of CH₂Cl₂. The organic solution was then dried over    magnesium sulfate, filtered and concentrated to yield 3.26 g of the    title product as a clear oil. (¹H) NMR (D₂O): δ=3.82 (s, 3H), 4.43    (dd, 1H), 4.53 (dd, 1h), 4.67 (t, 1H), 6.29 (s, 1H).-   B. (S)-4-Hydroxymethyl-oxazlidin-2-one. To a solution of 3.26 g of    the resultant compound of Example 109A in 20 mL of ethanol at 0° C.    was added 0.85 g of sodium borohydride in small portions. The ice    bath was removed and after additional 3 hours, 20 mL of 2.0 N    hydrogen chloride was added to the mixture, which was then    concentrated to yield an oil. The residue was extracted with EtOAc    and the organic solution was dried over magnesium sulfate, filtered    and concentrated to yield 2.50 g of the title compound. (¹H) NMR    (CDCl₃): δ=2.48 (s, 1H), 3.69 (dd, 1H), 4.08 (m, 1H), 4.31 (t, 1H),    4.57 (t, 1H).-   C. 4-Nitrophenyl-((S)-4-oxazlidin-2-onyl)-methyl carbonate. To a    solution of 1.04 g of p-nitrophenyl chloroformate in 20 mL of CH₂Cl₂    at 0° C. was added sequentially, 0.5 g of the resultant compound of    Example 109B and 0.6 mL of 4-methyl morpholine. The mixture was    stirred for 2 hours at ambient temperature and then concentrated in    vacuo. The residue was purified by low pressure silica gel column    chromatography using 20% EtOAc in CH₂Cl₂ eluent to yield 0.57 g of    the title compound. TLC: Rf=0.10, 50% EtOAc/hexane.-   D. Compound 109. To a solution of 60 mg of the resultant compound of    Example 35A in 1 mL of THF was added sequentially, 56 μL of    diisopropylethylamine and a solution of 51.1 mg of the resultant    compound of Example 109C in 1 mL acetonitrile. The mixture was    stirred for 24 hours and then concentrated in vacuo. The residue was    purified by preparative thin layer chromatography using 5%    methanol/CH₂Cl₂ as eluent to yield 60.4 mg of the title compound.    TLC: Rf=0.38, 5% methanol/CH₂Cl₂. HPLC: Rt=14.11 min. (¹H) NMR    (CDCl₃) consistent with structure.

EXAMPLE 110

Compound 110. To a solution of 60 mg of the resultant compound ofExample 40A in 1 mL of acetonitrile was added sequentially, 51 μL ofdiisopropylethylamine and a solution of 46.8 mg of the resultantcompound of Example 109C in 1 mL acetonitrile. The mixture was stirredfor 48 hours and then concentrated in vacuo. The residue was purified bypreparative thin layer chromatography using 10 methanol/CH₂Cl₂ eluentfollowed by preparative reversed-phase C18 HPLC using a linear gradientof 30% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to yield 16 mg of thetitle compound. TLC: Rf=0.28, 50% EtOAc/CH₂Cl₂. HPLC: Rt=12.47 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 111

A solution of 0.067 mmol of the resultant compound of Example 114D in 5mL of tetrahydrofuran was added 20 μL of diisopropylethylamine followeddropwise by a solution of the resultant compound of Example 82A in 5 mLof tetrahydrofuran during one hour. The mixture was stirred 16 h andthen concentrated in vacuo. The crude residue was purified by thicklayer silica gel chromatography using 5% MeOH/CH₂Cl₂ as eluant to obtain21.8 mg of the title compound. TLC: Rf=0.45, 5% MeOH/CH₂Cl₂; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 112

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,    E=3-sulfonamidophenyl). To a solution of 96.6 mg (0.287 mmol) of the    resultant compound of Example 39A in CH₂Cl₂ (4 mL) was added aqueous    sodium bicarbonate (1 mL), solid sodium bicarbonate 36.2 mg (0.431    mmol), and m-benzene disulfonylchloride 86.9 mg, (1.08 mmol). After    stirring for 1 h, 30% ammonium hydroxide (10 mL) was added. After 14    h the resulting mixture was diluted with CH₂Cl₂, washed with    saturated brine, dried over magnesium sulfate, filtered and    concentrated in vacuo. The residue was purified by flash    chromatography using 0% to 10% methanol/CH₂Cl₂ eluent to provide    49.3 mg of the title product. (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound XXII (A=H, D′=isobutyl, E=3-sulfonamidophenyl,    hydrochloride salt). A solution of 49.3 mg (0.089 mmol) of the    resultant compound of Example 112A in EtOAc (10 mL) at −20° C. was    treated with anhydrous HCl gas for 10 min. The ice bath was removed    and after an additional 15 min., the reaction mixture was sparged    with nitrogen then concentrated in vacuo to provide 53.1 mg of title    product as the HCl salt. (¹H)-NMR (CDCl₃) consistent with structure.-   C. Compound 112. To a solution of 53.1 mg of the resultant compound    of Example 112B (0.089 mmol) in CH₂Cl₂ (3 mL) was treated    sequentially at ambient temperature under an atmosphere of nitrogen,    with 0.031 mL (0.177 mmol) diisopropylethylamine and 24.3 mg (0.106    mmol) of the resultant compound of Example 48A. The mixture was    stirred 16 h and then concentrated in vacuo. The residue was taken    up in CH₂Cl₂ and washed with saturated brine, dried over magnesium    sulfate, filtered, and concentrated in vacuo. The residue was    purified by low pressure silica gel column chromatography using a    gradient 5% to 20% EtOAc in CH₂Cl₂ as eluent to yield 10.8 mg of the    title product. TLC: RF=0.4 25% EtOAc in CH₂Cl₂. HPLC: Rt=13.3 min;    (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 113

-   A. 3-Furansulfonyl chloride. In flame dried glassware under a    nitrogen atmosphere to a solution of 428 mg (2.909 mmol) of 3    bromofuran in anhydrous tetrahydrofuran at −78° C. was added 2.0 mL    n-butyl lithium (3.2 mmol at 1.6 molar in hexane). After 45 minutes    the resultant solution was added via cannula to a 20° C. solution of    sulfuryl chloride in diethyl ether (5 mL plus a 2 mL rinse). After 1    h, the reaction was quenched with 0.5 N hydrochloric acid and    extraced into diethyl ether. The ethereal extracts were washed with    saturated brine, dried over magnesium sulfate, filtered and    concentrated in vacuo to provide 158 mg of the title product. (¹H)    NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl, E=3-furyl). To    a solution of 289.7 mg (0.861 mmol) of the resultant compound of    Example 39A in CH₂Cl₂ (8 mL) was added aqueous sodium bicarbonate (2    mL), solid sodium bicarbonate 108 mg (1.292 mmol), and the resultant    product from Example 113A 157.8 mg, (1.08 mmol). After stirring for    1 h 30% ammonium hydroxide (10 mL) was added. After 14 h, the    resulting mixture was diluted with CH₂Cl₂, washed with saturated    brine, dried over magnesium sulfate, filtered and concentrated in    vacuo. The residue was purified by flash chromatography using 1% to    15% EtOAc/CH₂Cl₂.-   C. Compound XXII (A=H, D′=isobutyl, E=3-furyl, hydrochloride salt).    A solution of 217.3 mg (0.581 mmol) of the resultant compound of    Example 113B in EtOAc (15 mL) at −20° C. was treated with anhydrous    HCl gas for 10 min. The ice bath was removed and after an additional    15 min. the reaction mixture was sparged with nitrogen then    concentrated in vacuo to provide 228 mg of title product as the HCl    salt. TLC: Rf=0.52 10% methanol/CH₂Cl₂.-   D. Compound 113. To a solution of 65.3 mg of the resultant compound    of Example 113C (0.162 mmol) in CH₂Cl₂ (3 mL) was treated    sequentially at ambient temperature under an atmosphere of nitrogen,    with 0.056 mL (0.324 mmol) diisopropylethylamine and 44.6 mg (0.194    mmol) of the resultant compound of Example 48A. The mixture was    stirred 16 h and then concentrated in vacuo. The residue was taken    up in CH₂Cl₂ and washed with saturated brine, dried over magnesium    sulfate, filtered, and concentrated in vacuo. The residue was    purified by low pressure silica gel column chromatography using a    gradient 3% to 20% EtOAc in CH₂Cl₂ eluent to yield 10.8 mg of the    title product. TLC: Rf=0.6, 25% EtOAc/CH₂Cl₂. HPLC: Rt=13.9 min;    (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 114

-   A. Aminomethylcyclopentane. To a solution of LiAlH₄ (38 g, 1.0 mole)    in diethyl ether (2 L) was added cyclopentanecarbonitrile (73.2 g,    0.77 mol) as a solution in 250 mL ether. The solution was stirred    overnight at ambient temperature and then quenched by addition of    the organics to 3 L of a saturated potassium, sodium tartrate    solution. The amine was extracted into 3 L of ether, dried over    anhydrous K₂CO₃ then concentrated by distillation to approximately    400 mL total volume. The crude product was purified via distillation    to give 58.2 g of the title compound as a colorless oil. (¹H)-NMR    (CDCl₃) consistent with structure.-   B. Compound XXI (P=tert-butoxycarbonyl, D′=cyclopentylmethyl, P′=H).    To the resultant compound of Example 114A (20 g, 0.2 mol) was added    compound XX (P=Boc) (5.84 g) and the mixture was stirred for 24 h at    ambient temperature. The solution was concentrated by distillation    under reduced pressure. The residue was triturated with hexane and    the solid collected by suction filtration and washed with hexane to    give 7.08 g of a white solid which was used without further    purification. TLC: Rf=0.59 (1:10:90 concentrated    NH₄OH/methanol/CH₂Cl₂), (¹H)-NMR (CDCl₃) consistent with structure.-   C. Compound XXII (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=4-fluorophenyl). To a solution of the resultant compound of    Example 114B (200 mg, 0.55 mmol) in CH₂Cl₂ (10 mL) was added    4-fluorobenzenesulfonyl chloride (210 mg, 1.1 mmol) followed by the    addition of a saturated solution of sodium bicarbonate (3 mL)    followed by addition of solid sodium bicarbonate (0.1 g, 1.2 mmol).    The mixture was allowed to stir at ambient temperature overnight.    The solution was diluted with 100 mL CH₂Cl₂, the organics separated,    dried over anhydrous MgSO₄, and the organics concentrated under    reduced pressure to obtain 0.33 g of crude product. This material    was purified via medium pressure liquid chromatography using CH₂Cl₂,    followed by 0.5:99.5 methanol/CH₂Cl₂, followed by a 1:99    methanol/CH₂Cl₂ solution as the solvent system to give 120 mg (42%    yield) of the title compound as a white solid. TLC: Rf=0.48 (3:97    methanol/CH₂Cl₂); HPLC: Rt=18.22 min, (¹H)-NMR (CDCl₃) consistent    with structure.-   D. Compound XXII (P=H, D′=cyclopentylmethyl, E=4-fluorophenyl,    hydrochloride salt). A solution of 266 mg of the resultant compound    of Example 114C in ethyl acetate was treated at −20° C. with HCl gas    for 20 min, during which time the temperature was allowed to warm to    20° C. Nitrogen was then bubbled through the mixture for 15 min and    the solvent removed in vacuo to yield 224 mg of white solid which    was used directly for ensuing reaction.-   E. Compound 114. A solution of 31 mg of the resultant compound of    Example 114D in CH₂Cl₂ was treated sequentially, at ambient    temperature, under an atmosphere of nitrogen, with 9 mg of allyl    chloroformate and 19 mg N,N-diisopropylethylamine. The mixture was    stirred for 3 h and then concentrated in vacuo. The residue was    taken up in ethyl acetate and washed in 0.5 N HCl and saturated NaCl    then dried over MgSO₄, filtered, and concentrated in vacuo to yield    34 mg of the title product as a white solid. TLC: Rf=0.34 (5%    diethyl ether in CH₂Cl₂). HPLC: Rt=17.21 min. (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 115

Compound 115. A solution of 31 mg of the resultant compound of Example114B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of ethyl chloroformate and 19 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 35 mg of the title productas a white solid. TLC: Rf=0.32 (5% diethyl ether/CH₂Cl₂). HPLC: Rt=16.86min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 116

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=4-chlorophenyl). The resultant compound of Example 114B (252 mg)    was reacted with 4-chlorobenzenesulfonyl chloride (175 mg) in the    manner described in Example 166A. Workup and purification by silica    gel chromatography using EtOAc/CH₂Cl₂ as eluant yielded the product    as a white solid; (¹H) NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=H, D′=cyclopentylmethyl, E=4-chlorophenyl,    hydrochloride salt). A solution of 320 mg of the resultant compound    of Example 116A in 20 mL of EtOAc was treated with anhydrous HCl gas    for 5 min. The reaction mixture was sparged with nitrogen then    concentrated in vacuo to yeld a white solid which was used directly    for subsequent reaction.-   C. Compound 116. To a solution of 63.4 mg of the resultant compound    of Example 116B in 1 mL of THF was added sequentially 54 μL of    diisopropylethylamine and a solution of 39.9 mg of the resultant    compound of Example 48A in 1 mL THF. The mixture was stirred for 24    hours and then concentrated in vacuo. The residue was purified by    low pressure silica gel column chromatography using 20% EtOAc in    CH₂Cl₂ eluent to yield 0.62 g of the title compound. TLC: Rf=0.71,    40% EtOAc/CH₂Cl₂. HPLC: Rt=16.88 min. (¹H) NMR (CDCl₃) consistent    with structure.

EXAMPLE 117

Compound 117. A solution of 66.1 mg of the resultant compound of Example116B in 1 mL of THF was treated sequentially with 56 μL ofdiisopropylethylamine and 19.3 μL of allyl chloroformate. The mixturewas stirred for 4 hours and concentrated in vacuo. The residue was takeninto 50 mL of EtOAc and washed with 1.0 N HCl, saturated sodiumbicarbonate, brine, dried over magnesium sulfate, filtered andconcentrated. The residue was purified by low pressure silica gel columnchromatography using 20% EtOAc in hexane eluent to yield 69.7 mg of thetitle compound. TLC: Rf=0.20, 20% EtOAc/hexane. HPLC: Rt=17.83 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 118

Compound 118. To a solution of 65.3 mg of the resultant compound ofExample 116B in 1 mL of THF was added sequentially 55 μL ofdiisopropylethylamine and a solution of 49.2 mg of the resultantcompound of Example 82A in 1 mL THF. The mixture was stirred for 24hours and concentrated in vacuo. The residue was purified by lowpressure silica gel column chromatography using 40% EtOAc in CH₂Cl₂ aseluent followed by preparative reversed-phase C₁₈ HPLC using a lineargradient of 40% to 80% CH₃CN/H₂O for elution to yield 70.7 mg of thetitle compound. TLC: Rf=0.27, 40% EtOAc/CH₂Cl₂. HPLC: Rt=14.85 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 119

Compound 119. A solution of 26 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 6 mg of ethyl chloroformate and 15 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetage andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 26 mg of the title product as a white solid. TLC:Rf=0.19 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=17.50 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 120

Compound 120. A solution of 30 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of ethyl chloroformate and 18 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetage andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using ethyl acetate aseluent to yield 25 mg of the title product as a white solid. TLC:Rf=0.60 (ethyl acetate). HPLC: Rt=13.86 min. (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 121

Compound 121. A solution of 26 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 7 mg of ethyl chloroformate and 17 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 22 mg of the title product as a white solid. TLC:Rf=0.14 (5% diethyl ether/CH₂Cl₂). HPLC: Rt=15.95 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 122

Compound 122. A solution of 27 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of allyl chloroformate and 18 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether in CH₂Cl₂ aseluent to provide 23 mg of the title product as a white solid. TLC:Rf=0.33, 5% diethyl ether in CH₂Cl₂. HPLC: Rt=16.28 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 123

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,    E=3,4-dimethoxyphenyl). To a solution of 401 mg (1.192 mmol) of the    resultant compound of Example 39A in CH₂Cl₂ (12 mL) was added    aqueous sodium bicarbonate (3 mL), solid sodium bicarbonate 130 mg    (1.549 mmol), and 3,4-dimethoxybenzenesulfonyl chloride 33.8 mg,    (1.43 mmol). After 14 h, the resulting mixture was diluted with    EtOAc, washed with saturated brine, dried over magnesium sulfate,    filtered and concentrated in vacuo. The residue was purified by    flash chromatography using 5% to 25% EtOAc/CH₂Cl₂ eluent to provide    440.1 mg of the title product. TLC: Rf=0.72, 20% EtOAc/CH₂C₂.-   B. Compound XXII (A=H, D′=isobutyl, E=3,4-dimethoxyphenyl,    hydrochloride salt). A solution of 440 mg (0.820 mmol) of the    resultant compound of Example 123A in EtOAc (15 mL) at −20° C. was    treated with anhydrous HCl gas for 10 min. The ice bath was removed    and after an additional 15 min. the reaction mixture was sparged    with nitrogen then concentrated in vacuo to provide 610 mg of title    product as the HCl salt. TLC: Rf=0.44, 10% methanol/CH₂Cl₂.-   C. Compound 123. A solution of 38.9 mg of the resultant compound of    Example 123B (0.170 mmol) in CH₂Cl₂ (3 mL) was treated sequentially    at ambient temperature under an atmosphere of nitrogen with 0.049 mL    (0.283 mmol) diisopropylethylamine and 66.9 mg (169.6 mmol) of the    resultant compound of Example 48A. The mixture was stirred for 16 h    and then concentrated in vacuo. The residue was taken up in CH₂Cl₂    and washed with saturated brine, dried over magnesium sulfate,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel column chromatography using a gradient 10% to    25% diethyl ether in CH₂Cl₂ cluent to yield 57.6 mg of the title    product. TLC: Rf=0.39, 25% diethyl ether/CH₂Cl₂. HPLC: Rt=14.3 min;    (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 124

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl, E=3,4    difluorophenyl). To a solution of 332.7 mg (0.989 mmol) of the    resultant compound of Example 39A in CH₂Cl₂ (12 mL) was added    aqueous sodium bicarbonate (3 mL), solid sodium bicarbonate 125 mg    (1.483 mmol), and 3,4 difluorobenzensulfonyl chloride 231 mg. (1.088    mmol). After 14 h, the resulting mixture was diluted with CH₂Cl₂,    washed with saturated brine, dried over magnesium sulfate, filtered    and concentrated in vacuo. The residue was purified by flash    chromatography using 5% to 25% diethyl ether/CH₂Cl₂ eluent to    provide 313.6 mg of the title product. (¹H)-NMR (CDCl₃) consistent    with structure.-   B. Compound XXII (A=H, D′=isobutyl, E=3,4 difluorophenyl,    hydrochloride salt). A solution of 312.6 mg (0.610 mmol) of the    resultant compound of Example 124A in EtOAc (15 mL) at −20° C. was    treated with anhydrous HCl gas for 10 min. The ice bath was removed    and after an additional 15 min., the reaction mixture was sparged    with nitrogen then concentrated in vacuo to provide 280 mg of title    product as a white solid. TLC: Rf=0.46, 10% methanol/CH₂Cl₂.-   C. Compound 124. To a solution of 64.7 mg of the resultant compound    of Example 124B (0.144 mmol) in CH₂Cl₂ (3 mL) was treated    sequentially at ambient temperature under an atmosphere of nitrogen,    with 0.050 mL (0.288 mmol) diisopropylethylamine and 39.6 mg (172.9    mmol)of the resultant compound of Example 48A. The mixture was    stirred for 16 h and then concentrated in vacuo. The residue was    taken up in CH₂Cl₂ and washed with saturated brine, dried over    magnesium sulfate, filtered, and concentrated in vacuo. The residue    was purified by low pressure silica gel column chromatography using    a gradient 5% to 20% diethyl ether in CH₂Cl₂ eluent to yield 44 mg    of the title product. TLC: RF=0.54 25% diethyl ether/CH₂Cl₂. HPLC:    Rt=15.4 min. (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 125

Compound 125. This compound was prepared from the resultant compound ofExample 146B in the manner described in Example 88. After workup andpurification by preparative reversed-phase C₁₈ HPLC using a linergradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant, 10.5 mg ofthe title compound was obtained as a white solid. TLC: Rf=0.4, 4%MeOH/CH₂Cl₂. HPLC: Rt=14.06 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 126

-   A. Compound XXI (P=tert-butoxycarbonyl, D′=methyl, P′=H). To a    solution compound XX (1.7 mmol) in ethanol (20 mL) was added    methylamine gas, at ambient temperature, for 30 min. The solution    was stirred overnight, then concentrated under reduced pressure to    give 0.47 g of the title compound which was used without subsequent    purification. TLC: Rf=0.19, 1:10:90 NH₄OH/methanol/CH₂Cl₂, (¹H)-NMR    (CDCl₃) consistent with structure.-   B. Compound 126. To a solution of the product of Example 126A (0.15    g, 0.51 mmol) in CH₂Cl₂ (10 mL) was added a saturated solution of    sodium bicarbonate (3 mL), followed by addition of solid sodium    bicarbonate (90 mg, 1.1 mmol), followed-by addition of    3,4-dichlorobenzenesulfonyl chloride (0.25 g, 1.0 mmol). The mixture    was stirred at ambient temperature overnight. The organics were    extracted into 100 mL CH₂Cl₂, dried over anhydrous, MgSO₄,    concentrated under reduced pressure then purified via medium    pressure silica gel chromatography using a gradient system of CH₂Cl₂    followed by 5:95 ether/CH₂Cl₂. The title compound was obtained as a    colorless foam 210 mg. TLC: Rf=0.42 (3:97 methanol/CH₂Cl₂), HPLC:    Rt=17.2 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 127

Compound 127. To a solution of the product of Example 126A (0.15 g, 0.51mmol) in CH₂Cl₂ (10 mL) was added a saturated solution of sodiumbicarbonate (3 mL), followed by addition of solid sodium bicarbonate(100 mg, 1.0 mmol), followed by addition of 4-fluorobenzenesulfonylchloride (0.20 g, 1.0 mmol). The mixture was stirred at ambienttemperature overnight. The organics were extracted into 100 mL CH₂Cl₂,dried over anhydrous, MgSO₄, concentrated under reduced pressure thenpurified via medium pressure silica gel chromatography using a gradientsystem of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂. The title compound wasobtained as a white solid 104 mg. TLC: Rf=0.36, 3:97 methanol/CH₂Cl₂,HPLC: Rt=15.86 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 128

Compound 128. To a solution of the product of Example 126A (0.15 g, 0.51mmol) in CH₂Cl₂ (6 mL) was added a saturated solution of sodiumbicarbonate (3 mL), followed by addition of solid sodium bicarbonate (90mg, 1.0 mmol), followed by addition of acetamidobenzenesulfonyl chloride(0.24 g, 1.02 mmol). The mixture was stirred at ambient temperatureovernight. The organics were extracted into 100 mL CH₂Cl₂, dried overanhydrous, MgSO₄, concentrated under reduced pressure then purified viamedium pressure silica gel chromatography using a gradient system ofCH₂Cl₂ followed by 5:95 EtOAc/CH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂.The title compound was obtained as 244 mg of white solid. TLC: Rf=0.13,3:97 methanol/CH₂Cl₂, HPLC: Rt=13.47 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 129

-   A. Compound XXI (P=tert-butoxycarbonyl,    D′=(2-tetrahydrofuryl)-methyl, P′=H). To a solution compound XX (3.3    mmol) in ethanol (30 mL) was added tetrahydrofurfurylamine (1.03 mL,    10 mmol). The mixture was warmed to 85° C. and stirred overnight.    The solution was filtered and the solution concentrated under    reduced pressure to give 1.29 g of the title compound which was used    without subsequent purification. TLC: Rf=0.52, 1:10:90    NH₄OH/methanol/CH₂Cl₂-   B. Compound 129. To a solution of the resultant compound of Example    129A (200 mg, 0.55 mmol) in CH₂Cl₂ (6 mL) was added    4-fluorobenzenesulfonyl chloride (320 mg, 1.6 mmol) followed a    saturated solution of sodium bicarbonate (3 mL) and solid sodium    bicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambient    temperature overnight. The solution was diluted with 100 mL CH₂Cl₂,    the organics separated, dried over anhydrous MgSO₄. and the organics    concentrated under reduced pressure. The crude product was purified    via medium pressure liquid chromatography using a gradient solvent    system of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂ followed by a 10:90    ether/CH₂Cl₂ solution to give 130 mg of the title compound as a    white solid. TLC: Rf=0.35, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.37 min,    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 130

-   A. Compound XXI (P=tert-butoxycarbonyl, D′=(isobutenyl, P′=H)). To a    solution compound XX (P=tert-butoxycarbonyl) (2.5 mmol) in ethanol    (30 mL) was added a solution 2-methylallylamine hydrochloride (1.34    g, 12.5 mmol) and KOH (0.70 g, 12.5 mmol) in ethanol (20 mL). The    mixture stirred 30 min at ambient temperature. The solutions were    combined and heated to 85° C. for 24 h. The solution was filtered    and concentrated under reduced pressure to give 0.82 g of the title    compound which was used without subsequent purification. TLC:    Rf=0.45, 1:10:90 concentrated NH₄OH/methanol/CH₂Cl₂.-   B. Compound 130. To a solution of the product of Example 130A (0.20    g, 0.60 mmol) in CH₂Cl₂ (6 mL) was added a saturated solution of    sodium bicarbonate (3 mL), followed by solid sodium bicarbonate (0.1    g, 1.2 mmol) and then p-fluorobenzenesulfonyl chloride (0.35 g, 1.78    mmol). The mixture was stirred at ambient temperature for 24 h. The    organics were extracted into 100 mL CH₂Cl₂, dried over anhydrous    MgSO₄, concentrated under reduced pressure then purified via medium    pressure silica gel chromatography using a gradient system of    CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂. The title compound was    obtained as a white solid 180 mg. TLC: Rf=0.35, 3:97    methanol/CH₂Cl₂, HPLC: Rt=16.82 min; (¹H)-NMR (CDCl₃) consistent    with structure.

EXAMPLE 131

Compound 131. To a solution of the resultant compound of Example 130A(200 mg, 0.60 mmol) in CH₂Cl₂ (6 mL) was added4-acetamidobenzenesulfonyl chloride (410 mg, 1.76 mmol), followed by asaturated solution of sodium bicarbonate (3 mL) and solid sodiumbicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambienttemperature overnight. The solution was diluted with 100 mL CH₂Cl₂, theorganics separated, dried over anhydrous MgSO₄, and the organicsconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using a gradient solvent system ofCH₂Cl₂, followed by 30:70 EtOAc/CH₂Cl₂ solution to give 140 mg of thetitle compound as a white solid. TLC: Rf=0.19, 3:97 methanol/CH₂Cl₂,HPLC: Rt=15.06 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 132

-   A. Compound XXII (A=H, D′=(2-tetrahydrofuryl)-methyl,    E=4-fluorophenyl, hydrochloride salt). To a solution of the    resultant compound of Example 129B (30 mg, 0.057 mmol) in EtOAc (3    mL) was added 30% w/w HCl in EtOAc (1 mL). The mixture was stirred    overnight at ambient temperature. The solution was concentrated    under reduced pressure to give 16 mg of the title compound as a    white solid which was used without subsequent purification. TLC:    Rf=0.60 (1:10:90 NH₄OH/methanol/CH₂Cl₂).-   B. Compound 132. To a solution of the resultant compound of Example    132A (16 mg) in CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72    mmol) followed by the compound of Example 48A (20 mg, 0.09 mmol).    The mixture was stirred at ambient temperature for 24 hours. The    solution was concentrated under reduced pressure and the crude    product purified via medium pressure colum chromatography using    20:80 EtOAc/CH₂Cl₂ as the solvent system to give 7.4 mg. Rf=0.37    (3:97 methanol/CH₂Cl₂), HPLC: Rt=14.19 min, (¹H)NMR (CDCl₃)    consistent with structure.

EXAMPLE 133

-   A. Compound XXII (A=tert-butoxycarbonyl,    D′=(2-tetrahydrofuryl)-methyl, E=4-acetamidophenyl). To a solution    of the resultant compound of Example 129A (200 mg, 0.55 mmol) in    CH₂Cl₂ (6 mL) was added 4-acetamidobenzenesulfonyl chloride (380 mg,    1.6 mmol) followed by a saturated solution of sodium bicarbonate (3    mL) and solid sodium bicarbonate (0.1 g, 1.2 mmol). the mixture was    stirred at ambient temperature overnight. The solution was diluted    with 100 mL CH₂Cl₂, the organics separated, dried over anhydrous.    MgSO₄, and the organics concentrated under reduced pressure. The    crude product was purified via medium pressure liquid chromatography    using a gradient solvent system of CH₂Cl₂, followed by 10:90    EtOAc/CH₂Cl₂, followed by a 30:70 EtOAc/CH₂Cl₂ solution to give 120    mg of the title compound as a white solid. TLC: Rf=0.13, 3:97    methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=H, D′=(2-tetrahydrofuryl)-methyl,    E=4-acetamidophenyl, hydrochloride salt). To a solution of the    resultant compound of Example 133A (120 mg 0.22 mmol) in EtOAc (5    mL) was added 30% w/w HCl in EtOAc (2 mL). The mixture was stirred    overnight at ambient temperature. The solution was concentrated    under reduced pressure to give the title compound which was used    without subsequent purification. TLC: Rf=0.50, 1:10:90    NH₄OH/methanol/CH₂Cl₂.-   C. Compound 133. To a solution of the resultant compound of Example    133B in CH₂Cl₂ (5 mL) was added triethylamine (0.2 mL, 1.4 mmol)    followed by the compound of Example 48A (73 mg, 0.32 mmol). The    mixture was stirred at ambient temperature for 24 hours. The    solution was concentrated under reduced pressure and the crude    product purified via medium pressure column chromatography using a    gradient solvent system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,    followed by 3:97 methanol/CH₂Cl₂ as the solvent system to give 87.8    mg. Rf=0.09, 3:97 methanol/CH₂Cl₂, HPLC: Rt=12.53 min, (₁H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 134

-   A. Compound XXII (A=H, D′=isobutenyl, E=4-acetamidophenyl,    hydrochloride salt). To a solution of the resultant compound of    Example 131 (40 mg 0.075 mmol) in EtOAc (5 mL) was added 30% w/w HCl    in EtOAc (2 mL). The mixture was stirred overnight at ambient    temperature. The solution was concentrated under reduced pressure to    give the title compound, which was used without subsequent    purification. TLC: Rf=0.38, 1:10:90 NH₄OH/methanol/CH₂Cl₂.-   B. Compound 134. To a solution of the resultant compound of Example    134A in CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol),    followed by the compound of Example 48A (26 mg, 0.11 mmol). The    mixture was stirred at ambient temperature for 24 hours. The    solution was concentrated under reduced pressure and the crude    product purified via medium pressure column chromatography using a    gradient solvent system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,    followed by 3:97 methanol/CH₂Cl₂ as the solvent system to give 10.1    mg of the title compound. Rf=0.11 (3:97 methanol/CH₂Cl₂), HPLC:    Rt=12.86 min, (₁H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 135

-   A. Compound XXI (A=H, D′=(isobutenyl, E=4-fluoropheynl,    hydrochloride salt). To a solution of the resultant compound of    Example 130B (50 mg, 0.10 mmol) in EtOAc (5 mL) was added 30% w/w    HCl in EtOAc (1 mL). The mixture was stirred overnight at ambient    temperature. The solution was concentrated under reduced pressure to    give the title compound which was used without subsequent    purification. TLC: Rf=0.48, 1:10:90 NH₄OH/methanol/CH₂Cl₂.-   B. Compound 135. To a solution of the resultant compound of Example    135A in CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol),    followed by the compound of Example 48A (35 mg, 0.15 mmol). The    mixture was stirred at ambient temperature for 24 hours. The    solution was concentrated under reduced pressure and the crude    product purified via medium pressure column chromatography using a    gradient solvent system of CH₂Cl₂ followed by 20:80 EtOAc/CH₂Cl₂ as    the solvent system to give 12 mg. Rf=0.34, 3:97 methanol/CH₂Cl₂,    HPLC: Rt=14.64 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 136

-   A. Compound XXI (A=tert-butoxycarbonyl, D′=2-furfuryl, A′=H). To a    solution compound XX (2.5 mmol) in ethanol (30 mL) was added    furfurylamine (0.67 mL, 7.5 mmol) and the mixture was heated to    85° C. for 24 h. The solution was filtered and concentrated under    reduced pressure to give 0.80 g of the title compound which was used    without subsequent purification. TLC: Rf=0.38, 1:10:90 concentrated    NH₄OH/methanol/CH₂Cl₂.-   B. Compound XXII (A=tert-butoxycarbonyl, D′2-furyl,    E=4-fluorophenyl). To a solution of the product of Example 136A    (0.20 g, 0.60 mmol) in CH₂Cl₂ (6 mL) was added a saturated solution    of sodium bicarbonate (3 mL), followed by addition of solid sodium    bicarbonate (0.1 g, 1.2 mmol), then p-fluorobenzenesulfonyl chloride    (0.32 g, 1.6 mmol). The mixture was stirred at ambient temperature    for 24 h. The organics were extracted into 100 mL CH₂Cl₂, dried over    anhydrous MgSO₄, concentrated under reduced pressure, then purified    via medium pressure silica gel chromatography using a gradient    system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂. The title    compound was obtained as a white solid (86.1 mg). TLC: Rf=0.17, 3:97    methanol/CH₂Cl₂, HPLC: Rt=16.5 min; (₁H)-NMR (CDCl₃) consistent with    structure.-   C. Compound XXII (A=H, D′=2-furyl, E=4-fluorophenyl, hydrochloride    salt). To a solution of the resultant compound of Example 136B (16    mg, 0.031 mmol) in EtOAc (3 mL) was added 30% w/w HCl in EtOAc (1    mL). The mixture was stirred overnight at ambient temperature. The    solution was concentrated under reduced pressure to give the title    compound, which was used without subsequent purification. TLC:    Rf=0.48, 1:10:90 NH₄OH/methanol/CH₂Cl₂.-   D. Compound 136. To a solution of the resultant compound of Example    136C in CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol),    followed by the resultant compound of Example 48A (11 mg, 0.05    mmol). The mixture was stirred at ambient temperature for 24 hours.    The solution was concentrated under reduced pressure and the crude    product purified via medium pressure column chromatography using a    gradient solvent system of CH₂Cl₂ followed by 20:80 EtOAc/CH₂Cl₂ as    the solvent system to give 4.9 mg. TLC: Rf=0.28, (3:97    methanol/CH₂Cl₂, HPLC: Rt=14.57 min, (₁H)-NMR (CDCl₃) consistent    with structure.

EXAMPLE 137

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=2-furyl,    E=4-acetamidophenyl). To a solution of the resultant compound of    Example 136B (200 mg, 0.55 mmol) in CH₂Cl₂ (6 mL) was added    4-acetamidobenzenesulfonyl chloride (390 mg, 1.7 mmol) followed by    saturated solution of sodium bicarbonate (3 mL) and solid sodium    bicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambient    temperature overnight. The solution was diluted with 100 mL CH₂Cl₂,    the organics separated, dried over anhydrous. MgSO₄, and the    organics concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using a gradient    solvent system of CH₂Cl₂ followed by 10:90 EtOAc/CH₂Cl₂, followed by    a 30:70 EtOAc/CH₂Cl₂ solution to give 100 mg of the title compound    as a white solid. TLC: Rf=0.19, 3:97 methanol/CH₂Cl₂, (¹H)-NMR    (CDCl₃) consistent with structure.-   B. Compound XXII (A=H, D′=2-furyl, E=4-acetamidophenyl,    hydrochloride salt). To a solution of the resultant compount of    Example 137A (30 mg, 0.054 mmol) in EtOAc (3 mL) was added 30% w/w    HCl in EtOAc (1 mL). The mixture was stirred overnight at ambient    temperature. The solution was concentrated under reduced pressure to    give the title compound which was used without subsequent    purification. TLC: Rf=0.37 (1:10:90 NH₄OH/methanol/CH₂Cl₂).-   C. Compound 137. To a solution of the resultant compound of Example    137a in CH₂Cl₂ (5 mL) was Added triethylamine (0.1 mL, 0.72 mmol)    followed by the compound of Example 48A (19 mg, 0.083 mmol). The    mixture was stirred at ambient temperature for 24 hours. The    solution was concentrated under reduced pressure and the crude    product purified via medium pressure column chromatography using a    gradient solvent system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,    followed by 3:97 methanol/CH₂Cl₂ as the solvent system to give 8.5    mg of the title compound. TLC: Rf=0.11 (3:97 methanol/CH₂Cl₂), HPLC:    Rt=12.69 min; (₁H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 138

Compound 138. A solution of 75 mg of the resultant compound of Example51D and 45 mg of 3-chlorobenzenesulfonyl chloride were reacted in themanner described in Example 60. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 29.7 mg of the title compoundwas obtained. TLC: Rf=0.3, 4% MeOH/CH₂Cl₂, HPLC: Rt=15.83 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 139

Compound 139. To a solution of 67.9 mg of the resultant compount ofExample 116B in 1 mL of THF was added sequentially, 57 μL ofdiisopropylethylamine and a solution of 52.6 mg of the resultantcompound of Example 109C in 1 mL THF. The mixture was stirred for 24hours and concentrated in vacuo. The residue was purified by preparativethick layer silica gel chromatography using 7% methanol in CH₂Cl₂ eluentto yield 70.0 mg of the title compound. TLC: Rf=0.30, 5%methanol/CH₂Cl₂. HPLC: Rt=15.78 min; (¹H) NMR (CDCl₃) consistent withstructure.

EXAMPLE 140

-   A. 3(S)-amino-2(syn)-hydroxy-4-phenyl-1-chlorobutane formate salt.    To a slurry of 16.33 g of 10% palladium on carbon (25% by weight) in    methanol and tetrahydrofuran (400 mL, 1:1) was added, under N₂,    65.35 g of    3(S)-N-(-benzyloxycarbonyl)-amino-1-chloro-2(syn)-hydroxy-4-phenylbutane    (195.77 mmol) as a solution in methanol and tetrahydrofuran (1.2 L).    To this slurry was added 540 mL of formic acid. After 15 h, the    reaction mixture was filtered through diatomaceous earth and    concentrated to dryness. The resultant oil was slurried in toluene    and evaporated, then triturated sequentially with diethyl ether and    CH₂Cl₂ to provide 47.64 g of product as a granular tan solid. TLC:    Rf=0.17, 5% acetic acid/ethyl acetate.-   B.    3(S)-N-(3(S)-tetrahydrofuryloxycarbonyl)-amino-1-chloro-2(syn)-hydroxy-4-phenylbutane.    To a solution of the resultant compound of Example 140A (1.97 g,    7.95 mmol) in CH₂Cl₂ (20 mL) was added a saturated solution of    sodium bicarbonate (5 mL), followed by solid sodium bicarbonate    (1.33 g, 17.9 mmol), and the resultant compound of Example 48A (2.0    g, 8.7 mmol). The mixture was stirred at ambient temperature    overnight. The solution was diluted with 200 mL CH₂Cl₂, the organics    separated, dried over anhydrous MgSO₄, and concentrated under    reduced pressure. The residue was recrystallized from ethyl    acetate/hexane to give 1.01 g of the title compound as a white    solid. TLC: Rf=0.35, 3:97 methanol/CH₂Cl₂. (¹H)-NMR (CDCl₃)    consistent with structure.-   C. Compound XX (A=3(s)-tetrahydrofuryloxycarbonyl). To a solution of    the resultant compound of Example 140B (1.0 g, 3.2 mmol) in absolute    ethanol (15 mL) was added solid KOH (0.21 g, 3.8 mmol). The mixture    was stirred at ambient temperature for 1.0 h. The solution was    filtered through a pad of Celite then concentrated under reduced    pressure. The residue was taken up in ether (100 mL), washed with    brine, dried over MgSO₄, the concentrated under reduced pressure to    give 0.88 g of the title compound as a white solid. TLC: Rf=0.49    (3:97 methanol/CH₂Cl₂), (¹H)-NMR (CDCl₃) consistent with structure.-   D. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl,    D′=cyclopentylmethyl, A′=H). The resultant compound of Example 140C    (0.88 g, 3.2 mol) was added to the resultant compound of Example    114A (5.0 g, 50.4 mmol) and stirred for 24 h at ambient temperature.    The solution was concentrated by distillation under reduced    pressure. The residue was triturated with hexane and the solid    collected by suction filtration and washed with hexane to give 0.93    g of the title compound. TLC: Rf=0.44, 1:10:90 concentrated    NH₄OH/methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.-   E. Compound 140. To a solution of the resultant compound of Example    140D (0.93 g, 2.47 mmol) in CH₂Cl₂ (20 mL) was added a saturated    solution of sodium bicarbonate (5 mL) followed by addition of solid    sodium bicarbonate (0.42 g, 4.94 mmol) and 4-methoxybenzenesulfonyl    chloride (0.61 g, 2.96 mmol). the mixture was stirred at ambient    temperature for 4 hours. The solution was diluted with 200 mL    CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄, and the    organics concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using CH₂Cl₂    followed by 1:99 methanol/CH₂Cl₂ solution as the eluent system to    give 1.28 g of the title compound as a white solid. TLC: Rf=0.26,    3:97 methanol/CH₂Cl₂, HPLC: Rt=15.66 min, (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 141

-   A. Compound XXII (A=H, D′=cyclopentylmethyl, E=4-methoxyphenyl,    hydrochloride salt). A solution of 71.3 mg of the resultant compound    of Example 166A in EtOAc (25 mL) at 0° C. was treated with anhydrous    HCl gas for 10 min., and allowed to stand for 12 h while warming to    ambient temperature, then concentrated under reduced pressure and    the resulting white solid used without purification for subsequent    reaction.-   B. Compound 141. The resultant compound of Example 141A (0.134 mmol)    was reacted with allyl chloroformate in the manner described in    Example 87B. After concentration of the mixture in vacuo and workup,    the residue was purified by thick layer silica gel chromatography    using 5% MeOH/CH₂Cl₂ as eluant followed by preparative    reversed-phase C₁₈ HPLC using a linear gradient of 35% to 100%    CH₃CN/H₂O with 0.1% TFA as eluant to obtain 21.6 mg of the title    compound. TLC: Rf=0.45, 5% MeOH/CH₂Cl₂. HPLC: Rt=16.96 min.

EXAMPLE 142

Compound 142. To a solution of 4.0 g of the resultant compound ofExample 141A in 45 mL of THF was added sequentially, 1.96 mL ofdiisopropylethylamine and a solution of 2.68 g of the resultant compoundof Example 82A in 45 mL THF. The mixture was stirred for 24 hours andconcentrated in vacuo. The residue was taken up in CH₂Cl₂, washed withsaturated sodium bicarbonate and brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by lowpressure silica gel column chromatography using 20% to 40% EtOAc inhexane eluent to yield 3.69 g of the title compound. TLC: Rf=0.41, 50%EtOAc/CH₂Cl₂.

EXAMPLE 143

Compound 143. A solution of 3.69 g of the resultant compound of Example142 in 100 mL of ethyl ether was treated with anhydrous HCl gas for 10min. The reaction mixture was sparged with nitrogen then filtered. Thesolid was taken up in methanol and concentrated to yield 3.71 g of thetitle compound. TLC: Rf=0.62, 90/10/1 CH₂Cl₂/MeOH/AcOH, HPLC: Rt=13.87min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 145

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,    E=2-(5-isoxazoy-3-yl)-thiophene). To a solution of 342.5 mg (1.02    mmol) of the resultant compound of Example 39A in CH₂Cl₂ (8 mL) was    added aqueous sodium bicarbonate (2 mL), solid sodium bicarbonate    257 mg (3.1 mmol), and 5-(isoxazol-3-yl)-thiophenesulfonyl chloride    254.2 mg, (1.02 mmol). After 14 h, the resulting mixture was diluted    with CH₂Cl₂, washed with saturated brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by flash chromatography using 5% to 25% EtOAc/CH₂Cl₂ eluent    and recrystallized from ether CH₂Cl₂ to provide 228.6 mg of the    title product. (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A-H, D′=isobutyl, E=2-(5-isoxazoy-3-ly)-thiopene,    hydrochloride salt). A solution of 228.6 mg (0.416 mmol) of the    resultant compound of Example 145A in EtOAc (15 mL) at −20° C. was    treated with anhydrous HCl gas for 10 min. The ice bath was removed    and after an additional 15 min, the reaction mixture was sparged    with nitrogen then concentrated in vacuo to provide 223.6 mg of    title product as the HCl salt. TLC: Rf=0.48, 10% methanol/CH₂Cl₂.-   C. Compound 145. A solution of 78.5 mg of the resultant compound of    Example 145B (0.162 mmol) in CH₂Cl₂ (3 mL) was treated sequentially    at ambient temperature under an atmosphere of nitrogen with 0.07 mL    (0.408 mmol) diisopropylethylamine and 55.6 mg (0.243 mmol) of the    resultant compound of Example 48A. The mixture was stirred 16 h and    then concentrated in vacuo. The residue was taken up in CH₂Cl₂ and    washed with saturated brine, dried over magnesium sulfate, filtered,    and concentrated in vacuo. The residue was purified by preparative    HPLC to yield 48.7 mg of the title product. TLC: Rf=0.36, 25%    EtAOc/CH₂Cl₂. HPLC: Rt=15.2 min; (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 146

-   A. Compound XXI (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=4-acetamidophenyl). To a solution of the resultant compound of    Example 114B (300 mg, 0.83 mmol) in CH₂Cl₂ (15 mL) was added    4-acetamidobenzenesulfonyl chloride (580 mg, 2.48 mmol) followed by    the addition of a saturated solution of sodium bicarbonate (4 mL)    and solid sodium bicarbonate (0.14 g, 1.67 mmol). The mixture was    stirred at ambient temperature overnight. The solution was diluted    with 150 mL CH₂Cl₂, the organics separated, dried over anhydrous.    MgSO₄, and the organics concentrated under reduced pressure. The    crude product was purified via medium pressure liquid chromatography    using a gradient solvent system of CH₂Cl₂, followed by 5:95    EtOAc/CH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂ solution to give 310 mg    of the title compound as a white solid. TLC: Rf=0.10, 3:97    methanol/CH₂Cl₂, HPLC: Rt=15.96 min, (¹H)-NMR (CDCl₃) consistent    with structure.-   B. Compound XXII (P=H, D′=cyclopentylmethyl, E=4-acetamidophenyl,    hydrochloride salt). To a solution of the resultant compound of    Example 146A (210 mg, 0.38 mmol) was added 30% w/w HCl in EtOAc (15    mL). The mixture was stirred for 1 hour at ambient temperature. The    solution was concentrated under reduced pressure to give 180 mg of    the title compound which was used without subsequent purification.    TLC: Rf=0.14, 1:10:90 NH₄OH/methanol/CH₂Cl₂.-   C. Compound XXII (P=allyloxycarbonyl, D′=cyclopentylmethyl,    E=4-acetamidophenyl). To a solution of the resultant compound of    Example 146B (100 mg, 0.20 mmol) in CH₂Cl₂ (10 mL) was added    triethylamine (0.1 mL, 0.72 mmol), followed by allylchloroformate    (0.04 mL, 0.3 mmol). The mixture was stirred at ambient temperature    for 24 hours. The solution was diluted with 150 mL CH₂Cl₂, washed    with water, dried over anhydrous MgSO₄, and the organics    concentrated under reduced pressure. The crude product was purified    via medium pressure column chromatography using a gradient solvent    system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂, followed by 3:97    methanol/CH₂Cl₂ as the solvent system to give 103 mg. Rf=0.22, 3:97    methanol/CH₂Cl₂, HPLC: Rt=15.29 min, (¹H)-NMR (CDCl₃) consistent    with structure.

EXAMPLE 147

Compound 147. To a solution of the resultant compound of Example 146B(80 mg, 0.16 mmol) in CH₂Cl₂(5 mL) was added triethylamine (0.07 mL,0.48 mmol), followed by slow addition over 3 hours of the resultantcompound of Example 82A (53 mg, 0.19 mmol) as a solution in CH₂Cl₂ (3mL). The mixture was stirred at ambient temperature for 24 hours. Thesolution was diluted with 100 mL CH₂Cl₂ washed with water, dried overanhydrous MgSO₄, and the organics concentrated under reduced pressure.The crude product was purified via medium pressure column chromatographyusing a gradient solvent system of CH₂C₂, followed by 1:99methanol/CH₂Cl₂, followed by 2:98 methanol/CH₂Cl₂ as the solvent systemto give 71.7 mg of the title compound. Rf=0.06, 3:97 methanol/CH₂Cl₂,HPLC: Rt=12.61 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 148

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=phenyl). A solution of 297 mg of the resultant compound of Example    114B in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 217 mg of benzenesulfonyl chloride and 103 mg of    sodium bicarbonate. The mixture was stirred for 6 h, diluted with    CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄, filtered;    and concentrated in vacuo to yield 426 mg of the title product as a    white solid. TLC: Rf=0.32, 5% diethyl ether/CH₂Cl₂. (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (A=H, D′=cyclopentylmethyl, E=phenyl, hydrochloride    salt). A solution of 400 mg of the resultant compound of Example    148A in ethyl acetate was treated at −20° C. with HCl gas for 20    min, during which time the temperature was allowed to warm to 20° C.    Nitrogen was then bubbled through the mixture for 15 min and solvent    removed in vacuo to yield 349 mg of white solid which was used    directly for the ensuing reaction.-   C. Compound 148. A solution of 40 mg of the resultant compound of    Example 148B in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 31 mg of the resultant    compound of Example 48A and 35 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 14 h, diluted with CH₂Cl₂,    washed with saturated NaHCO₃ and saturated NaCl, then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was purified    by low pressure silica gel chromatography using 20% diethyl    ether/CH₂Cl₂ as eluent to provide 45 mg of the title product as a    white solid. TLC: Rf=0.46, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=15.78    min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 149

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentymethyl,    E=3-pyridyl). To a solution of 153 mg (0.422 mmol) of the resultant    compound of Example 114B in CH₂Cl₂ (4 mL) was added aqueous sodium    bicarbonate (1 mL), solid sodium bicarbonate 141.7 mg (1.69 mmol),    and the resultant compound of Example 144A 156.1 mg. (0.879 mmol).    After 14 h, the resulting mixture was diluted with CH₂Cl₂, washed    with saturated brine, dried over magnesium sulfate, filtered and    concentrated in vacuo. The residue was purified by flash    chromatography using 20% to 40% EtOAc/CH₂Cl₂ eluent to provide 64.7    mg of the title product. TLC; Rf=0.24, 20% EtOAc/CH₂Cl₂.-   B. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=3-pyridyl, hydrochloride salt). A solution of 273.1 mg (0.572    mmol) of the resultant compound of Example 149A in EtOAc (15 mL) at    −20° C. was treated with anhydrous HCl gas for 10 min. The ice bath    was removed and after an additional 15 min., the reaction mixture    was sparged with nitrogen then concentrated in vacuo. To a solution    of the resulting residue in CH₂Cl₂ (3 mL) was added, sequentially at    ambient temperature under an atmosphere of nitrogen, with 0.076 mL    (0.437 mmol) diisopropylethylamine and 34.3 mg (0.150 mmol) of the    resultant compound of Example 48A. The mixture was stirred for 16 h    and then concentrated in vacuo. The residue was taken up in CH₂Cl₂    and washed with saturated brine, dried over magnesium sulfate,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel column chromatography using a gradient 20% to    50% EtOAc in CH₂Cl₂ eluent to yield 11.3 mg of the title product.    TLC; Rf=0.15 40% EtOAc/CH₂Cl₂. HPLC: Rt=13.7 min; (¹H) NMR (CDCl₃)    consistent with structure.

EXAMPLE 150

-   A. 1-Piperidinesulfonyl chloride. A solution of 4 g of sulfuryl    chloride in acetronitrile was treated dropwise with 861 mg of    piperidine at ambient temperature under an atmosphere of nitrogen.    After complete addition, the mixture was refluxed for 18 h, cooled    to room temperature and concentrated in vacuo to yield the title    product as a red oil. TLC: Rf=0.86, CH₂Cl₂. (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,    E=piperidinyl). A solution of 73 mg of the resultant compound of    Example 39A in CH₂Cl₂ was treated sequentially, at ambient    temperature under an atmosphere of nitrogen, with 121 mg of the    resultant compound of Example 150A and 84 mg of    N,N-diisopropylethylamine. The mixture was stirred for 14 h, diluted    with CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ as    eluent to provide 70 mg of the title product as a white solid. TLC:    Rf=0.21 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=17.40 min. (¹H)-NMR    (CDCl₃) consistent with structure.-   C. Compound XXII (A=H, D′=isobutyl, E=piperidinyl, hydrochloride    salt). A solution of 70 mg of the resultant compound of Example 150B    in ethyl acetate was treated at −20° C. with HCl gas for 20 min    during which time the temperature was allowed to warm to 20° C.    Nitrogen was then bubbled through the mixture for 15 min and solvent    removed in vacuo to yield a viscous oil which was used directly for    the ensuing reaction.-   D. Compound 150. A solution of the resultant compound of Example    150C in CH₂Cl₂ was added, at ambient temperature under an atmosphere    of nitrogen, to a solution of 50 mg of the resultant compound of    Example 48A and 56 mg N,N-diisopropylethylamine in CH₂Cl₂. the    mixture was stirred for 14 h, diluted with CH₂Cl₂, washed with    saturated NaHCO₃ and saturated NaCl, then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel chromatography using 20% diethyl ether/CH₂Cl₂ as    eluent to provide 16 mg of the title product as a white solid. TLC:    Rf=0.45, (0% diethyl ether/CH₂Cl₂. HPLC: Rt=15.00 min. (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 151

-   A. Compound XXII (A=tert butoxycarbonyl, D′=cyclopentylmethyl,    E=4-trifluormethoxyphenyl). A solution of 71 mg of the resultant    compound of Example 114B in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was    treated sequentially, at ambient temperature under an atmosphere of    nitrogen, with 76 mg of 4-trifluoromethoxybenzensulfonyl chloride    and 25 mg of sodium bicarbonate. The mixture was stirred, 14 h,    diluted with CH₂Cl₂, washed with saturated NaCl then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was purified    by low pressure silica gel chromatography using 5% diethyl    ether/CH₂Cl₂ as eluent to provide 92 mg of the title product as a    white solid. TLC: Rf=0.34, 5% diethyl ether/CH₂Cl₂. (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (A=H, D′=cyclopentylmethyl,    E=4-trifluormethoxyphenyl, hydrochloride salt). A solution of 92 mg    of the resultant compound of Example 151A in ethyl acetate was    treated at −20° C. with HCl gas for 20 min, during which time the    temperature was allowed to warm to 20° C. Nitrogen was then bubbled    through the mixture for 15 min and solvent removed in vacuo to yield    83 mg of white solid which was used directly for the ensuing    reaction.-   C. Compound 151. A solution of 22 mg of the resultant compound of    Example 151B in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 15 mg of the resultant    compound of Example 48A and 16 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 60 h, diluted with CH₂Cl₂,    washed with saturated NaHCO₃ and saturated NaCl, then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was purified    by low pressure silica gel chromatography using 20% diethyl    ether/CH₂Cl₂ as eluent to provide 23 mg of the title product as a    white solid. TLC: Rf=0.44, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=16.99    min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 152

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,    E=4-trifluormethoxyphenyl). A solution of 97 mg of the resultant    compound of Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was    treated sequentially, at ambient temperature under an atmosphere of    nitrogen, with 113 mg of 4-trifluoromethoxybenzenesulfonyl chloride    and 36 mg of sodium bicarbonate. The mixture was stirred for 14 h,    diluted with CH₂Cl₂, washed with saturated NaCl then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was purified    by low pressure silica gel chromatography using 5% diethyl    ether/CH₂Cl₂ as eluent to provide 120 mg of the title product as a    white solid. TLC: Rf=0.34, 5% diethyl ether/CH₂Cl₂. HPLC: Rt=18.54    min. (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=H, D′=isobutyl, E=4-trifluormethoxyphenyl,    hydrochloride salt). A solution of 100 mg of the resultant compound    of Example 152A in ethyl acetate was treated at −20° C. with HCl gas    for 20 min, during which time the temperature was allowed to warm to    20° C. Nitrogen was then bubbled through the mixture for 15 min and    solvent removed in vacuo to yield 89 mg of white solid which was    used directly for ensuing reaction.-   C. Compound 152. A solution of 41 mg of the resultant compound of    Example 152B in CH₂Cl₂ was added, at ambient temperature under the    atmosphere of nigrogen, to a solution of 28 mg of the resultant    compound of Example 48A and 32 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred 14 h, diluted with CH₂Cl₂, washed    with saturated NaHCO₃ and saturated NaCl, then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel chromathography using 5% diethyl ether/CH₂Cl₂ as    eluent to provide 30 mg of the title product as white solid. TLC: Rf    −0.08 (5% diethyl ether/CH₂Cl₂). HPLC: Rt=16.52 min. (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 153

-   A. Compound XXII (A=tert-butoxycarbonyl,D′=isobutyl,    E=4-methoxyphenyl). To a solution of the resultant compound of    Example 39A (600 mg, 1.77 mmol) in CH₂Cl₂ (10 mL) was added    4-methoxybenzenesulfonyl chloride (0.55 g. 2.66 mmol) followed by    the addition of a saturated solution of sodium bicarbonate (3 mL)    and 0.30 g of solid sodium bicarbonate. The mixture was stirred at    ambient temperature overnight. The solution was diluted with 200 mL    CH₂Cl₂, the organics were separated, dried over anhydrous MgSO₄, and    the organics concentrated under reduced pressure. The crude product    was purified via medium pressure liquid chromatography using a    gradient solvent system of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂    solution to give 630 mg of the title compound as a white solid. TLC:    Rf=0.48, 3:97 methanol/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound XXII (A=H, D′=isobutyl, E=4-methoxyphenyl, hydrochloride    salt). To a solution of the resultant compound of Example 153A (0.63    g, 1.24 mmol) in EtAc (5 mL) was added 30% w/w HCl in EtOAc (5 mL.)    The mixture was stirred for 6 hours ambient temperature. The    solution was concentrated under reduced pressure to give 0.59 g of a    white solid which was used directly for subsequent reaction. TLC    Rf=0.12, 3:97 methanol/CH₂Cl₂.-   C. Compound XXII (A=(3-pyridyl)-methyloxycarbonyl, D′=isobutyl,    E=4-methoxyphenyl). To a solution of the resultant compound of    Example 153B (100 mg, 0.23 mmol) in CH₂Cl₂ (5 mL) was added    triethylamine (0.1 mL, 0.72 mmol) followed by slow addition over 3    hours of the resultant compound of Example 82A (75 mg, 0.27 mmol) as    a solution in CH₂Cl₂ (5 mL). The mixture was stirred at ambient    temperature for 24 hours. The organics was concentrated under    reduced pressure and the crude product was purified via medium    pressure column chromatography using a gradient solvent system of    CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂, followed by 3:97    methanol/CH₂Cl₂ as the solvent system to give 49.3 mg of the title    compound. Rf=0.33, 3:97 methanol/CH₂Cl₂. HPLC: Rt=13.18 min,    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 154

Compound 154. To a solution of the resultant compound of Example 153B(100 mg, 0.20 mmol) in CH₂Cl₂ (5 mL) was added triethylamine (0.25 mL,1.8 mmol) followed by allylchlorofornate (0.1 mL, 0.94 mmol). Themixture was stirred at ambient temperature for 24 hours. The solutionwas concentrated under reduced pressure and the crude product purifiedvia medium pressure column chromatography using a gradient solventsystem of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂ as the solvent systemto give 94 mg of the title compound. Rf=0.71, 3:97 methanol/CH₂Cl₂.HPLC: Rt=16.12 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 155

-   A. N-hydroxysuccinimidyl-1-methoxypropane-3-carbonate. A solution of    355 mg of 2-methylene-1,3-propanediol in acetonitrile (30 mL) was    added sequentially, at ambient temperature, 65 mg of sodium hydride    and 0.25 mL iodomethane. The mixture was stirred for 12 h and    concentrated in vacuo. The residue was then taken up in 15 mL of    acetonitrile and treated sequentially, at ambient temperature under    an atmosphere of nitrogen, with 1.3 g of N,N-disuccinimidyl    carbonate and 1.6 mL of triethylamine. After stirring for 14 h, the    reaction mixture was concentrated in vacuo and the residue was    diluted CH₂Cl₂, washed with saturated sodium bicarbonate solution    and saturated brine, dried over magnesium sulfate, filtered and    concentrated in vacuo. The residue was purified by silica gel    chromatography with EtOAc as eluant to give 95 mg of the title    compound. (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound 155. A solution of 0.056 mmol of the resultant compound    of Example 40A was reacted with the resultant compound of Example    155A in the manner described in Example 132. After concentration of    the mixture in vacuo and workup, the residue was purified by thick    layer silica gel chromatography using 7% MeOH/CH₂Cl₂ as eluant    followed by preparative reversed-phase C₁₈ HPLC using a linear    gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain    3.7 mg of the title compound. TLC: Rf=0.45, 7% MeOH/CH₂Cl₂. HPLC:    Rt=13.78 min.

EXAMPLE 156

-   A. 1-acetylindoline-5-sulfonyl chloride. A 1.02 g portion of    1-acetylindoline was treated with 2 mL of chlorosulfonic acid at    0° C. The mixture was heated at 60° C. for 2 h, then treated with    crushed ice, filtered and dried to give 1.3 g of the title compound    which was used directly for subsequent reaction. TLC: Rf=0.18, 50%    EtOAc/hexane. (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=5-(N-acetyl)-indoline). To a solution of 60 mg of the resultant    compound of Example 114B in 15 mL of CH₂Cl₂ was added (5 mL)    saturated aqueous sodium bicarbonate solution, 50.0 mg sodium    bicarbonate, and 60 mg of the resultant compound of Example 156A.    After 4 h, the resulting mixture was diluted with CH₂Cl₂, washed    with saturated brine, dried over magnesium sulfate and filtered. The    mixture was then concentrated in vacuo to give the desired product    which was used directly for subsequent reastion. (¹H)-NMR (CDCl₃)    consistent with structure.-   C. Compound 156. A solution of 37 mg of the resultant compound of    Example 156B in EtOAc (15 mL) at 0° C. was treated with anhydrous    hydrogen chloride gas for 10 min., and allowed to stand for 12 h    while warming to ambient temperature. This crude material was then    reacted with allyl chloroformate in the manner described in Example    87B. After concentration of the mixture in vacuo and workup, the    residue was purified by thick layer silica gel chromatography using    7% MeOH/CH₂Cl₂ as eluant followed by preparative reversed-phase C₁₈    HPLC using a linear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA    as eluant to obtain 10.5 mg of the title compound. TLC: Rf=0.75, 10%    MeOH/CH₂Cl₂. HPLC: Rt=15.78 min; (¹H)-NMR (CDCL₃) consistent with    structure.

EXAMPLE 157

Compound 157. A solution of 37 mg of the resultant compound of Example156B in EtOAc (15 mL) at 0° C. was treated with anhydrous hydrogenchloride gas for 10 min., and allowed to stand for 12 h while warming toambient temperature. This crude material was then reacted with theresultant compound of Example 48A in the manner described in Example 88.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain 17.9 mg of the titlecompound. TLC: Rf=0.6, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.68 min; (¹H)-NMR(CDCL₃) consistent with structure.

EXAMPLE 158

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl, E=H).    To a solution of compound XX (A=Boc) (5.0 mmol) in ethanol (20 mL)    was added cyclohexylmethylamine (3.25 mL, 2.83 mmol) and the mixture    was stirred for 3 hours at ambient temperature. The solution was    filtered and concentrated under reduced pressure to give 1.49 g of a    white solid which was used directly for subsequent reaction. TLC:    Rf=0.14, 3:97 methanol/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with    structure.-   B. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl,    E=4-methoxyphenyl). To a solution of the resultant compound of    Example 158A (400 mg, 1.06 mmol) in CH₂Cl₂ (10 mL) was added    4-methoxybenzenesulfonyl chloride (0.66 g, 3.1 mmol) followed by    addition of a saturated solution of sodium bicarbonate (3 mL) and    0.18 g of solid sodium bicarbonate. The mixture was stirred at    ambient temperature overnight. The solution was diluted with 200 mL    CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄, and the    organics concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using CH₂Cl₂,    followed by 1:99 methanol/CH₂Cl₂ as the solvent system to give 340    mg of the title compound as a white solid. TLC: Rf=0.39, 3:97    methanol/CH₂Cl₂, (¹H)-NMR (CDCL₃) consistent with structure.-   C. Compound XXI (A=H, D′=cyclohexylmethyl, E=4-methoxyphenyl,    hydrochloride salt). To a solution of the resultant compound of    Example 158B (0.34 g, 0.62 mmol) in EtOAc (10 mL) was added 30% w/w    HCl in EtOAc (5 mL). The mixture was stirred for 3 hours at ambient    temperature. The solution was concentrated under reduced pressure to    give 0.3 g of a white solid which was used directly for subsequent    reaction. TLC: Rf=0.12, 3:97 methanol/CH₂Cl₂.-   D. Compound 158. To a solution of the resultant compound of Example    158C (100 mg, 0.21 mmol) in CH₂Cl₂ (8 mL) was added triethylamine    (0.2 mL, 1.44 mmol) followed by the resultant compound of Example    48A (71 mg, 0.31 mmol). The mixture was stirred at ambient    temperature for 6 hours. The solution was diluted with CH₂Cl₂, (200    mL) washed with a saturated solution of sodium bicarbonate (30 mL),    the organics separated, dried over anhydrous MgSO₄ and concentrated    under reduced pressure and the crude product purified via medium    pressure column chromatography using a gradient solvent system of    CH₂Cl₂ followed by 10:90 EtOAc/CH₂Cl₂ as the solvent system to give    84.9 mg of the title compound. TLC: Rf=0.48, 3:97 methanol/CH₂Cl₂,    HPLC: Rt=16.35 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 159

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl,    E=4-fluorophenyl). To a solution of the resultant compound of    Example 158A (400 mg, 1.06 mmol) in CH₂Cl₂ (10 mL) was added    4-fluorobenzenesulfonyl chloride (0.62 g, 3.2 mmol) followed by    addition of a saturated solution of sodium bicarbonate (3 mL) and    0.18 g of solid sodium bicarbonate. The mixture was stirred at    ambient temperature overnight. The solution was diluted with 200 mL    CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄, and the    organics concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using CH₂Cl₂    followed by 1:99 methanol/CH₂Cl₂ solution as the solvent system to    give 280 mg of a white solid. TLC: Rf=0.47, 3:97 methanol/CH₂Cl₂,    (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=H, D′=cyclohexylmethyl, E=4-fluorophenyl,    hydrochloride salt). To a solution of the resultant compound of    Example 159A (0.28 g, 0.52 mmol) was added 30% w/w HCl in EtOAc (10    mL). The mixture was stirred for 3 hours at ambient temperature. The    solution was concentrated under reduced pressure to give 0.23 g of a    white solid which was used directly for subsequent reaction. TLC:    Rf=0.13, (3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCL₃) consistent with    structure.-   C. Compound 159. To a solution of the resultant compound of Example    159C (100 mg, 0.21 mmol) in CH₂Cl₂ (8 mL) was added triethylamine    (0.2 mL, 1.44 mmol) followed by the resultant compound of Example    48A (73 mg, 0.32 mmol). The mixture was stirred at ambient    temperature for 6 hours. The solution was diluted with CH₂Cl₂, (200    mL) washed with saturated solution of sodium bicarbonate (30 mL),    dried over anhydrous MgSO₄, the organics concentrated under reduced    pressure and the crude product purified via medium pressure column    chromatography using a gradient solvent system of CH₂Cl₂, followed    by 10:90 EtOAc/CH₂Cl₂ as the solvent system to give 54 mg of the    title compound. TLC: Rf=0.46, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.48    min; (¹H)-NMR (CDCL₃) consistent with structure.

EXAMPLE 160

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl,    E=4-acetamidophenyl). To a solution of the resultant compound of    Example 158A (400 mg, 1.06 mmol) in CH₂Cl₂ (10 mL) was added    4-acetamidobenzenesulfonyl chloride (0.75 g, 3.2 mmol) followed by    addition of a saturated solution of sodium bicarbonate (3 mL) and    0.18 g of solid sodium bicarbonate. The mixture was stirred at    ambient temperature overnight. The solution was diluted with 200 mL    CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄, and the    organics concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using CH₂Cl₂,    followed by 1:99 methanol/CH₂Cl₂ and 2:98 methanol/CH₂Cl₂ as the    solvent system to give 290 mg of the title compound as a white    solid. TLC: Rf=0.14, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (A=H, D′=cyclohexylmethyl, E=4-acetamidophenyl,    hydrochloride salt). To the resultant compound of Example 160A (0.29    g, 0.51 mmol) was added 30% w/w HCl in EtOAc (10 mL). The mixture    was stirred for 3 hours at ambient temperature. The solution was    concentrated under reduced pressure to give 0.28 g of a white solid    which was used directly for subsequent reaction. TLC: Rf=0.10, 3:97    methanol/CH₂Cl₂.-   C. Compound 160. To a solution of the resultant compound of Example    160B (100 mg, 0.20 mmol) in CH₂Cl₂ (8 mL) was added triethylamine    (0.2 mL, 1.44 mmol) followed by the resultant compound of Example    48A (67 mg, 0.30 mmol). The mixture was stirred at ambient    temperature for 6 hours. The solution was diluted with CH₂Cl₂, (200    mL) washed with saturated solution of sodium bicarbonate (30 mL),    dried over anhydrous MgSO₄, the organics concentrated under reduced    pressure and the crude product purified via medium pressure column    chromatography using a gradient solvent system of CH₂Cl₂, followed    by 10:90 EtOAc/CH₂Cl₂, followed by 20:80 EtOAc/CH₂Cl₂ as the solvent    system to give 56.8 mg of a white solid. TLC: Rf=0.17, 3:97    methanol/CH₂Cl₂, HPLC: Rt=14.65 min; (¹H)-NMR (CDCl₃) consistent    with structure.

EXAMPLE 161

-   A. 4-Morpholinesulfonyl chloride. A solution of 4.6 g of sulfuryl    chloride in acetonitrile was treated dropwise with 996 mg of    morpholine at ambient temperature under an atmosphere of nitrogen.    After complete addition, the mixture was refluxed for 16 h, cooled    to room temperature, and concentrated in vacuo to yield the title    product as a red oil. TLC: Rf=0.65 CH₂Cl₂. (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (A-tert-butoxycarbonyl, D′=isobutyl,    E=morpholinyl). A solution of 98 mg of the resultant compound of    Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 270 mg of the resultant compound of Example 161A and    122 mg of sodium bicarbonate. The mixture was stirred for 14 h,    diluted with CH₂Cl₂, dried over MgSO₄, filtered, and concentrated in    vacuo. The residue was purified by low pressure silica gel    chromatography using CH₂Cl₂ as eluent followed by preparative HPLC    to provide 22 mg of the title product as an oily solid. TLC:    Rf=0.46, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=15.50 min. (¹H)-NMR    (CDCl₃) consistent with structure.-   C. Compound XXII (A=H, D′=isobutyl, E=morpholinyl, hydrochloride    salt). A solution of 22 mg of the resultant compound of Example 161B    in ethyl acetate was treated at −20° C. Nitrogen was then bubbled    through the mixture for 15 min and solvent removed in vacuo to yield    an oily semi-solid mass which was used directly for the ensuing    reaction.-   D. Compound 161. A solution of the resultant compound of Example    161C in CH₂Cl₂ was added, at ambient temperature under an atmosphere    of nitrogen, to a solution of 16 mg of the resultant compound of    Example 48A and 18 mg N,N-diisopropylethylamine in CH₂Cl₂. The    mixture was stirred for 14 h, diluted with CH₂Cl₂, washed and    saturated with NaHCO₃ and saturated NaCl, then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by    preparative HPLC to provide 21 mg of the title product as an oily    solid. TLC: Rf=0.22, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=13.01 min.    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 162

Compound 162. A solution of 30 mg of the resultant compound of Example166A was deprotected with hydrogen chloride gas and the resultantcompound was reacted with the resultant compound of Example 155A in themanner described in Example 155B. After concentration of the mixture invacuo and workup, the residue was purified by thick layer silica gelchromatography using 5% MeOH/CH₂Cl₂ as eluant, followed by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100%CH₃CN/H₂O) with 0.1% TFA as eluant to obtain 6.2 mg of the titlecompound. TLC: Rf=0.65, 5% MeOH/CH₂Cl₂. HPLC: Rt=15.93 min (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 163

Compound 163. A 120.3 mg portion of the resultant compound of Example153B was reacted with the resultant compound of Example 82A as describedin Example 82B. After workup and concentration in vacuo, the residue waspurified by low pressure silica gel column chromatography using 50%EtOAc in CH₂Cl₂ eluent, followed by preparative reversed-phase C₁₈ HPLCusing a linear gradient of 40% to 100% acetonitrile/water for elution toobtain 44.3 mg of the title compound. TLC: Rf=0.18, 50% EtOAc/CH₂Cl₂.HPLC: Rt=13.13 min; (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 164

-   A. N-hydroxysuccinimidyl-(2-phenyl)ethyl carbonate. A solution of    306 mg of phenethyl alcohol and 535 mg of N,N′-disuccinimidyl    carbonate in acetonitrile was treated, at ambient temperature under    an atmosphere of nitrogen, with 810 mg of N,N-diisopropylethylamine.    The mixture was stirred for 60 h. and concentrated in vacuo. The    residue was taken up in ethyl acetate and washed with saturated    NaHCO₃, saturated NaCl, then dried over MgSO₄, filtered, and    concentrated in vacuo to yield the title product as a yellow oil.    TLC: Rf=0.40 (5% methanol in CH₂Cl₂). (¹H)-NMR (CDCl₃) consistent    with structure.-   B. Compound 164. A solution of 81 mg of the resultant compound of    Example 164A in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 41 mg of the resultant    compound of example 40a and 45 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 4 h, diluted with CH₂Cl₂, washed    with saturated NaHCO₃ and saturated NaCl, then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was subjected to    preparative HPLC to yield 18 mg of the title product. TLC: Rf=0.83    (5:10:85 NH₄H/CH₃OH/CH₂Cl₂). HPLC: Rt=15.78 min. (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 165

Compound 165. A solution of 36 mg of the resultant compound of Example51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 20 mg ofp-toluenesulfonyl chloride and 18 mg of sodium bicarbonate. The mixturewas stirred for 3 h, diluted with CH₂Cl₂, washed with saturated NaClthen dried over MgSO₄, filtered, and concentrated in vacuo. The residuewas purified by low pressure silica gel chromatography using 5% diethylether/CH₂Cl₂ as eluent to provide 38 mg of the title product as a whitesolid. TLC: Rf=0.15, 5% diethyl ether/CH₂Cl₂. HPLC: Rt=15.27 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 166

-   A. Compound XXII (P=tert-butoxycarbonyl. D′=cyclopentylmethyl,    E=4-methoxyphenyl). To a solution of the resultant compound of    Example 114B (1.8 g, 4.96 mmol) in CH₂Cl₂ (10 mL) was added    4-methoxylbenzensulfonyl chloride (2.10 g, 9.93 mmol), followed by    addition of a saturated solution of sodium bicarbonate (3 mL) and    0.83 g of solid sodium bicarbonate. The mixture was stirred at    ambient temperature for 24 hours. The solution was diluted with 200    mL CH₂Cl₂, the organics were separated, dried over anhydrous MgSO₄,    and concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using CH₂Cl₂,    followed by 1:99 methanol/CH₂Cl₂ followed by 2:98 methanol/CH₂Cl₂ as    the solvent system to give 1.49 g of the title compound as a white    solid. TLC: Rf=0.37, 3:97 methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃)    consistent with structure.-   B. Compound XXII (P=H, D′=cyclopentylmethyl, E=4-hydroxyphenyl). A    solution of the resultant compound of Example 166A (1.11 g, 2.08    mmol) in CH₂Cl₂ (20 mL) was added to a solution of boron tribromide    in CH₂Cl₂ (1.0 M, 10.4 mL). The mixture was stirred at ambient    temperature for 24 hours. The solution was poured onto 40 mL of a    saturated solution of sodium bicarbonate. The aqueous layer was    extracted with 250 mL CH₂Cl₂ followed by extraction with 250 mL    EtOAc. The combined organics were dried over anhydrous MgSO₄,    concentrated under reduced pressure and the crude product purified    via medium pressure column chromatography using a gradient solvent    system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂, followed by 9:98    methanol/CH₂Cl₂, followed by a 1:5:95 concentrated    NH₄OH/methanol/CH₂Cl₂ solution as the solvent system to give 0.38 g    of the title compound. TLC: Rf=0.18, 3:97 methanol/CH₂Cl₂, (¹H)-NMR    (CDCl₃) consistent with structure.-   C. Compound 166. To a solution of the resultant compound of Example    166B (300 mg, 0.69 mmol) in CH₂Cl₂ (5 mL) was added triethylamine    (0.12 mL, 8.6 mmol), followed by slow addition over 3 hours of the    resultant compound of Example 82A (0.21 g, 0.77 mmol) as a solution    in CH₂Cl₂ (5 mL). The mixture was stirred at ambient temperature for    24 hours. The solution was diluted with 250 mL CH₂Cl₂, washed with    water, dried over anhydrous MgSO₄, and the organics concentrated    under reduced pressure. The crude product was purified via medium    pressure column chromatography using a gradient solvent system of    CH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂, followed by 2:98    methanol/CH₂Cl₂ as the solvent system to give 110 mg of a white    solid. TLC: Rf=0.14 (3:97 methanol/CH₂Cl₂), HPLC: Rt=12.69 min,    (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 167

Compound 167. A solution of 102 mg of the resultant compound of Example51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 65 mg ofp-nitrobenzenesulfonyl chloride and 51 mg of sodium bicarbonate. Themixture was stirred for 14 h, diluted with CH₂Cl₂, washed with saturatedNaCl, then dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was purified by low pressure silica gel chromatography using 20%diethyl ether/CH₂Cl₂ as eluent to provide 124 mg of the title product asa white solid. TLC: Rf=0.36, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=15.15min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 168

Compound 168. A solution of 124 mg of the resultant compound of Example167 in ethyl acetate was treated, at ambient temperature, with 13 mg of10% palladium on carbon. The mixture was stirred for 14 h under anatmosphere of hydrogen, filtered through a pad of Celite filter agent,and concentrated in vacuo. The residue was subjected to preparative HPLCto yield 82 mg of the title product as a white solid. TLC: Rf=0.10, 20%ether/CH₂Cl₂. HPLC: Rt=13.16 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 169

Compound 169. To a solution of the resultant compound of Example 166B(80 mg, 0.18 mmol) in CH₂Cl₂ (15 mL) was added a saturated solution ofsodium bicarbonate (5 mL) followed by the addition of the resultantcompound of Example 48A (55 mg, 0.24 mmol). The mixture was stirred atambient temperature for 5 hours. The solution was diluted with 200 mLCH₂Cl₂, the organics separated, dried over anhydrous MgSO₄, andconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂ as the solvent system to give 56 mg of the titlecompound as a white solid. TLC: Rf=0.24, 3:97 methanol/CH₂Cl₂, HPLC:Rt=14.29 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 170

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=4,nitrophenyl). To a solution of the resultant compound of Example    114B (250 mg, 0.69 mmol) in CH₂Cl₂ (15 mL) was added a saturated    solution of sodium bicarbonate (5 mL) followed by solid sodium    bicarbonate (0.12 g, 1.37 mmol) and 4-nitrobenzensulfonyl chloride    (200 mg, 0.9 mmol). The mixture was stirred at ambient temperature    for 24 hours. The solution was diluted with 200 mL CH₂Cl₂, the    organics separated, dried over anhydrous MgSO₄, and concentrated    under reduced pressure. The crude product was purified via medium    pressure liquid chromatography using a gradient solvent system of    CH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂ to give 360 mg of the title    compound as an orange solid. TLC: Rf=0.45, 3:97 methanol/CH₂Cl₂.    (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=H, D′=cyclopentylmethyl, E-4-nitrophenyl,    hydrochloride salt). To the resultant compound of Example 170A (360    mg, 0.66 mmol) was added 10% w/w HCl in EtOAc (15 mL). The mixture    was stirred for 3 hours at ambient temperature. The solution was    concentrated under reduced pressure to give 310 mg of the title    compound as an orange solid which was used directly for subsequent    reaction. TLC: Rf=0.70, 1:10:90 NH₄OH/methanol/CH₂Cl₂.-   C. Compound 170. To a solution of the resultant compound of Example    170B (310 mg, 0.64 mmol) in CH₂Cl₂ (15 mL) was added a saturated    solution of sodium bicarbonate (5 mL) followed by the addition of    solid sodium bicarbonate (0.11 g, 1.3 mmol) and the resultant    compound of Example 48A (0.18 g, 0.77 mmol). The mixture was stirred    at ambient temperature for 24 hours. The solution was diluted with    150 mL CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄,    and concentrated under reduced pressure. The crude product was    purified via medium pressure liquid chromatography using CH₂Cl₂,    followed by 1:99 methanol/CH₂Cl₂ as the solvent system to give 0.32    g of the title compound as a white solid. TLC: Rf=0.28, 3:97    methanol/CH₂Cl₂, HPLC: Rt=16.06 min, (¹H)-NMR (CDCl₃) consistent    with structure.

EXAMPLE 171

Compound 171. A solution of the resultant compound of Example 170C (0.19g, 0.34 mmol) in EtOAc (10 mL) was treated at ambient temperature with50 mg of 10% palladium on carbon and hydrogenated for 72 hours under aslight positive pressure of hydrogen. The mixture was filtered andconcentrated in vacuo and the crude product purified via medium pressureliquid chromatography using CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,followed by 3:97 methanol/CH₂Cl₂, followed by 10:90 methanol/CH₂Cl₂ asthe solvent system to give 97 mg of the title compound as a white solid.TLC: Rf=0.25, 3:97 methanol/CH₂Cl₂, HPLC: Rt=14.28 min, (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 172

-   A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,    E=2,4-dinitrophenyl). To a solution of the resultant compound of    Example 114B (500 mg, 1.38 mmol) in CH₂Cl₂ (15 mL) was added a    saturated solution of sodium bicarbonate (5 mL) followed by solid    sodium bicarbonate (0.23 g, 2.76 mmol) and    2,4-dinitrobenzenesulfonyl chloride (440 mg, 1.65 mmol). The mixture    was stirred at ambient temperature for 2 hours. The solution was    diluted with 200 mL CH₂Cl₂, the organics separated, dried over    anhydrous MgSO₄, and concentrated under reduced pressure. The crude    product was purified via medium pressure liquid chromatography using    a gradient solvent system of CH₂Cl₂, followed by 1:99    methanol/CH₂Cl₂ to give 700 mg of the title compound as a brown    solid. TLC: Rf=0.48, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃),    consistent with structure.-   B. Compound XXII (A=H, D′=cyclopentylmethyl, E-2,4-dinitrophenyl,    hydrochloride salt). To a the resultant compound of Example 172A    (700 mg, 1.18 mmol) was added 10% w/w HCl in EtOAc (20 mL). The    mixture was stirred for 3 hours at ambient temperature. The solution    was concentrated under reduced pressure to give 590 mg of the title    compound as a brown solid which was used without subsequent    purification. TLC: Rf=0.55, 1:10:90 NH₄OH/methanol/CH₂Cl₂.-   C. Compound 172. To a solution of the resultant compound of 172B    (590 mg, 1.11 mmol) in CH₂Cl₂ (15 mL) was added a saturated solution    of sodium bicarbonate (5 mL), followed by solid sodium bicarbonate    (0.19 g, 2.2 mmol) and the resultant compound of Example 48A (0.31    g, 1.3 mmol). The mixture was stirred at ambient temperature for 24    hours. The solution was diluted with 150 mL CH₂Cl₂, the organics    separated, dried over anhydrous MgSO₄, and the organics concentrated    under reduced pressure. The crude product was purified via medium    pressure liquid chromatography using a CH₃OH/CH₂Cl₂ gradient as    eluant, to yield the product as 0.59 g of a white solid. HPLC:    Rt=16.36 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 173

Compound 173. A solution of the resultant compound of Example 172C (0.20g, 0.33 mmol) in EtOAc (10 mL) was treated under ambient temperaturewith 50 mg of 10% palladium on carbon and hydrogenated for 72 hoursunder a slight positive pressure of hydrogen. The mixture was filteredand concentrated in vacuo and the crude product purified via mediumpressure liquid chromatography using CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂, 3:97 methanol/CH₂Cl₂, and 10:90 methanol/CH₂Cl₂ as thesolvent system to give 120.2 mg of the title compound as a light brownsolid. TLC: Rf=0.17, 3:97 methanol/CH₂Cl₂, HPLC: Rt=13.47 min, (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 174

-   A. 4-Benzyloxybenzenesulfonyl chloride. To 0.87 g of    dimethylformamide, at 0° C. under an atmosphere of nitrogen, was    added 1.61 g of sulfuryl chloride. The mixture was stirred for 15    min and treated with 2.00 g of benzyl phenyl ether. The mixture was    then heated at 100° C. for 1.5 h, cooled to about 40° C., poured    onto ice, extracted with CH₂Cl₂, dried over MgSO₄, filtered, and    concentrated in vacuo. The residue was purified by low pressure    silica gel chromatography using 10% ethyl acetate in hexane as    eluent to provide 0.78 g of the title product as a white solid. TLC:    Rf=0.46, 10% ethyl acetate in hexane. (¹H)-NMR (CDCl₃) consistent    with structure.-   B. Compound 174. A solution of 30 mg of the resultant compound of    Example 51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 24 mg of the resultant compound of Example 174A and    18 mg of sodium bicarbonate. The mixture was stirred for 14 h,    diluted with CH₂Cl₂, washed with saturated NaCl then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was purified    by low pressure silica gel chromatography using 20% diethyl    ether/CH₂Cl₂ as the eluent to provide 14 mg of the title product as    a white solid. TLC: Rf=0.43, 20% diethyl ether/CH₂Cl₂. HPLC:    Rt=17.01 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 175

Compound 175. A solution of 11 mg of the resultant compound Example 174Bin ethyl acetate was treated at ambient temperature, with 2 mg of 10%palladium on carbon. The mixture was stirred for 14 h under anatmosphere of hydrogen, filtered through a pad of Celite filter agent,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 10% methanol in CH₂Cl₂ as theeluent to provide 9 mg of the title product as a white solid. TLC:Rf=0.38, 10% methanol in CH₂Cl₂. HPLC: Rt=13.37 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 176

-   A. 1,3-Benzodioxole-5-sulfonyl chloride. To 3.50 g of    dimethylformamide, at 0° C. under an atmosphere of nitrogen, was    added 6.47 g of sulfuryl chloride. The mixture was stirred 15 min    and treated with 5.32 g of 1,3-benxodioxole. The mixture was then    heated at 120° C. for 45 min, cooled to about 40° C., poured onto    ice, extracted with CH₂Cl₂, dried over MgSO₄, filtered, and    concentrated in vacuo. The residue was purified by low pressure    silica gel chromatography using 40% CH₂Cl₂ in hexane as eluent to    provide 2.70 g of the title product as a yellow solid. TLC: Rf=0.37,    40% CH₂Cl₂ in hexane. (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound XXII (A=tert-butoxy, D′=isobutyl, E=3,4-benzodioxole). A    solution of 49 mg of the resultant compound of Example 39A in 4:1    CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, at ambient    temperature under an atmosphere of nitrogen, with 45 mg of the    resultant compound of Example 176A and 28 mg of sodium bicarbonate.    The mixture was stirred for 14 h, diluted with CH₂Cl₂, washed with    saturated NaCl then dried over MgSO₄, filtered, and concentrated in    vacuo. The residue was purified by low pressure silica gel    chromatography using 20% diethyl ether/CH₂Cl₂ as the eluent to    provide 71 mg of the title product as a waxy solid. TLC: Rf=0.65,    20% diethyl ether/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with    structure.-   C. Compound XXII (A=H, D′=isobutyl, E=3,4-benzodioxole,    hydrochloride salt). A solution of 71 mg of the resultant compound    of Example 176B in ethyl acetate was treated at −20° C. with HCl    gas. The HCl was bubbled through the mixture for 20 min over which    time the temperature was allowed to warm to 20° C. Nitrogen was then    bubbled through the mixture for 15 min and solvent removed in vacuo    to yield 66 mg of the title product as a white solid which was used    directly in subsequent reactions.-   D. Compound 176. A solution of 18 mg of the resultant compound of    Example 176C in CH₂Cl₂ was added, at ambient temperature under an    atmosphere of nitrogen, to a solution of 13 mg of the resultant    compound of Example 48A and 14 mg N,N-diisopropylethylamine in    CH₂Cl₂. The mixture was stirred for 16 h, diluted with CH₂Cl₂,    washed with saturated NaHCO₃ and saturated NaCl, then dried over    MgSO₄, filtered, and concentrated in vacuo. The residue was purified    by low pressure silica gel chromatography using 5% diethyl    ether/CH₂Cl₂ as the eluent to provide 9 mg of the title product as a    white solid. TLC: Rf=0.14, 5% diethyl ether CH₂Cl₂. HPLC: Rt=15.52    min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 177

-   A. (4-Methoxyphenyl)-methyl-4-nitrophenyl carbonate. To a solution    of 1.50 g of p-nitrophenyl chloroformate in 30 mL of CH₂Cl₂ at 0° C.    was added sequentially, 0.77 mL of 4-methoxybenzyl alcohol and 0.82    mL of 4-methyl morpholine. After stirring for a half hour at ambient    temperature, the resulting mixture was diluted with CH₂Cl₂, washed    with water, brine, dried over magnesium sulfate, filtered and    concentrated in vacuo to yield a pale yellow solid which was    triturated with CH₂Cl₂/hexane and filtered to yield 1.51 g of the    title compound. TLC: Rf=0.40, 20% EtOAc/hexane.-   B. Compound 177. To a solution of 96.7 mg of the resultant compound    of Example 141A in 2 mL of CH₂Cl₂ was added sequentially, 90 μL of    diisopropylethylamine and 81.3 mg of the resultant compound of    Example 178A. After stirring for 24 hours, the mixture was diluted    with CH₂Cl₂, washed with water and brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by preparative thin layer chromatography using 5% methanol    in CH₂Cl₂ eluent to yield 104.8 mg of the title compound. TLC:    Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.66 min, (¹H)NMR (CDCl₃)    consistent with structure.

EXAMPLE 178

-   A. (3-Methoxyphenyl)-methyl-4-nitrophenyl carbonate. Prepared by the    same route as described for Example 177A, except 3-methoxybenzyl    alcohol was utilized for reaction with p-nitrophenyl chloroformate    to yield the title compound as a pale yellow solid. TLC: Rf=0.40,    20% EtOAc/hexane.-   B. Compound 178. To a solution of 97.8 mg of the resultant compound    of Example 141A in 2 mL of CH₂Cl₂ was added sequentially, 91 μL of    disopropylethylamine and 82.2 mg of the resultant compound of    Example 178A. After stirring for 24 hours, the mixture was diluted    with CH₂Cl₂, washed with water and brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by preparative thin layer chromatography using 5% methanol    in CH₂Cl₂ eluent to yield 25.7 mg of the title compound. TLC:    Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.75 min. (¹H)NMR (CDCl₃)    consistent with structure.

EXAMPLE 179

-   A. (2-Methoxyphenyl)-methyl-4-nitrophenyl carbonate. Prepared by the    same route as described for Example 177A, except 2-methoxybenzyl    alcohol was utilized for reaction with p-nitrophenyl chloroformate    to yield the title compound as a pale yellow solid. TLC: Rf=0.40,    20% EtOAc/hexane.-   B. Compound 179. To a solution of 97.8 mg of the resultant compound    of Example 141A in 2 mL of CH₂Cl₂ was added sequentially, 99 μL of    diisoprophylethylamine and 89.2 mg of the resultant compound of    Example 179A. After stirring for 24 hours the mixture was diluted    with CH₂Cl₂, washed with water and brine, dried over magnesium    sulfate, filtered and concentrated in vacuo. The residue was    purified by preparative thin layer chromatography using 5% methanol    in CH₂Cl₂ eluent to yield 107.0 mg of the title compound. TLC:    Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.58 min. (¹H)NMR (CDCl₃)    consistent with structure.

EXAMPLE 180

-   A. 2,3-Dihydrobenzofuran-5-sulfonyl chloride. To. 3.35 g of    dimethylformamide, at 0° C. under an atmosphere of nitrogen, added    6.18 g of sulfuryl chloride. The mixture was stirred 15 min and    treated with 4.69 g of 2,3-dihydrobenzofuran. The mixture was then    heated at 100° C. for 1.5 h, cooled to about 40° C., poured onto    ice, extracted with CH₂Cl₂, dried over MgSO₄, filtered, and    concentrated in vacuo. The residue was taken up in ethyl acetate,    cooled to 5° C. for 16 h, and the resultant pink crystals collected    by vacuum filtration to provide 6.12 g of the title product. TLC:    Rf=0.41, 10% ethyl acetate in hexane. (¹H)-NMR (CDCl₃) consistent    with structure.-   B. Compound 180. A solution of 32 mg of the resultant compound of    Example 140D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 22 mg of the resultant compound of Example 180A and    18 mg of sodium bicarbonate. The mixture was stirred 14 h, diluted    with CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄,    filtered, and concentrated in vacuo. The residue was purified by low    pressure silica gel chromatography using 20% diethyl ether/CH₂Cl₂ as    eluent to provide 20 mg of the title product as a white solid. TLC:    Rf=0.52, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=15.49 min (¹H)-NMR    (CDCl₃) consistent with-structure.

EXAMPLE 181

Compound 181. A solution of the resultant compound of Example 140D (150mg, 0.4 mmol) in CH₂Cl₂ (10 mL) was added a saturated solution of sodiumbicarbonate (5 mL) followed by solid sodium bicarbonate (0.1 g, 1.2mmol) and 4-cyanobenzensulfonyl chloride (0.1 g, 0.48 mmol). The mixturewas stirred at ambient temperature for 4 hours. The solution was dilutedwith 200 mL CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄,and the organics concentrated under reduced pressure. The crude productwas purified via medium pressure liquid chromatography using CH₂Cl₂,followed by 1:99 methanol/CH₂Cl₂ solution as the solvent system to give0.19 g (86% yield) of the title compound as a white solid. TLC: Rf=0.40,3:97 methanol/CH₂Cl₂, HPLC: Rt=15.02 min, (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 182

Compound 182. This compound was prepared from the resultant compound ofExample 114D and the resultant compound of Example 48A in the samemanner described in Example 88. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 32.8 mg of the title compoundwas obtained. TLC: Rf=0.25, 4% MeOH/CH₂Cl₂. HPLC: Rt=16.06 min; (¹H)NMR(CDCl₃) consistent with structure.

EXAMPLE 183

Compound 183. This compound was prepared from the resultant compound ofExample 84 by treatment with hydrogen chloride gas and subsequentreaction with the resultant compound of Example 48A in the mannerdescribed in Example 132. After workup and purification bycrystallization from EtOAc, 33.0 mg of the title compound was obtainedas a white solid. TLC: Rf=0.25, 4% MeOH/CH₂Cl₂. HPLC: Rt=17.71 min;(¹H)-NMR (CDCL₃) consistent with structure.

EXAMPLE 184

-   A. (N-tert-butoxycarbonyl)-(R)-3-pyrrolidinyl-N-hydroxysuccinimidyl    carbonate. To a solution of 1.0 g of (R)-3-hydroxypyrrolidine in    tetrahydrofuran (50 mL) was added sequentially, at ambient    temperature, 3.75 g of di-tert-butyl dicarboante and 1 mL of 2N    sodium hydroxide. The mixture was stirred for 1 hour, filtered and    concentrated in vacuo. The resultant compound was reacted with    N,N-disuccinimidyl carbonate in the manner described in Example    155A. Workup and purification by thick layer silica gel    chromatography using an EtOAc eluent yielded the title compound as a    white solid; (¹H)-NMR (CDCl₃) consistent with structure.-   B. Compound 184. A solution of 350 mg of the resultant compound of    Example 166A was deprotected with hydrogen chloride gas and the    resultant compound was reacted with the resultant compound of    Example 184A in the manner described in Example 88. After    concentration of the mixture in vacuo and workup, the residue was    purified by thick layer silica gel chromatography using 7%    MeOH/CH₂Cl₂ as eluant, to obtain 120 mg of the title compound. TLC:    Rf=0.45, 5% MeOH/CH₂Cl₂. HPLC: Rt=16.97 min; (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 185

Compound 185. A solution of 120 mg of the resultant compound of Example184B in EtOAc (25 mL) at 0° C. was treated with anhydrous hydrogenchloride gas for 10 min., and allowed to stand for 12 h while warming toambient temperature. Concentration in vacuo yielded 110 mg of the titlecompound. TLC: Rf=0.35, 10% MeOH/89% CH₂Cl₂/1% NH₄OH. HPLC: Rt=13.72min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 186

-   A. Compound XXX ((syn, anti)-OH, A=carbobenzyloxy, R³=(s)-sec-butyl,    R^(3′)=H, D′=benzyl, A′=tert-butoxycarbonyl). A solution of 1.37 g    of the resultant compound of Example 1B in 150 mL of methylene    chloride was treated with 1.03 g of Cbz-Ile, 523 mg of HOBT.H₂O, and    742 mg of EDC. The mixture was stirred for 18 h, then diluted with 3    volumes of diethyl ether and washed sequentially with water,    saturated NaHCO₃ solution, 10% KHSO₄ solution, and brine. After    drying over MgSO₄ and concentrating in vacuo, the residue was    purified by chromatography on a silica gel column using a gradient    of 1% to 1.5% MeOH in CH₂Cl₂ as eluant to yield 2.10 g of the title    compound as a white foam. TLC: Rf=0.51, 5% methanol/CH₂Cl₂.-   B. Compound XXX ((syn, anti)-OH, A=carbobenzyloxy, R³=(S)-sec-butyl,    R^(3′)=H, D′=benzyl, A′=H), hydrochloride salt. A solution of 650 mg    of the resultant compound of Example 12A in 12 mL of ethyl acetate    was cooled in an ice/water bath and treated with a slow stream of    HCl gas for approximately 6 min with vigorous stirring. The mixture    was capped and stirred for an additional 10 min, then purged with a    stream of nitrogen for 15 minutes and concentrated in vacuo to yield    a white solid which was used without subsequent purification. TLC:    Rf=0.18, 95:5:0.5 CH₂Cl₂/methanol/concentrated NH₄OH.-   C. Compound 186. A solution of 20 mg of the resultant compound of    Example 186B in 0.8 mL of methylene chloride was cooled in    ice/methanol (approximately 15° C.), then treated with 13.8 μL of    DIEA followed by 7.6 mg of α-toluene sulfonyl chloride. The mixture    was stirred for 15 h, warming slowly to ambient temperature. The    mixture was concentrated to a small volume, applied to a 0.5 mm    thick prep plate and eluted with 3.5% MeOH/CH₂Cl₂. The band    containing the desired diastereomer was isolated and eluted with 8%    MeOH/CH₂Cl₂ to yield 4.8 mg of the title compound. TLC: Rf=0.42, 15%    diethyl ether/CH₂Cl₂. HPLC: Rt=17.81 min. NMR (CDCl₃): 0.78 (dd, 6H)    0.84 (m, 1H) 1.07, (m, 1H) 1.76-1.86 (m, 2H) 2.72 (m, 2H); 3.14 (s,    2H); 3.49 (dd, 1H); 3.87 (dd, 1H); 3.58 (m, 1H); 4.01 (d, 1H); 4.14,    (d, 1H); 4.26, (d, 1H); 4.35, (d, 1H); 4.90, (m, 1H); 5.08, (s, 2H);    5.97, (d, 1H), 7.08, (d, 2H); 7.17, (t, 1H); 7.20-7.40, (m, 17H).

EXAMPLE 187

Compound 187. 100 mg of the resulting compound 54A was treated with 1 mLof 90% aqueous TFA and allowed to stand for 12 h. The mixture wasconcentrated in vacuo and the residue taken up in 10 mL of dry CH₂Cl₂,treated with 65 mg of N-Cbz-L-isoleucine (0.235 mmol), 50 μL of DIEA(0.27 mmoles), 30 mg of HOBt (0.22 mmoles), and 42 mg of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.22mmoles). The mixture was stirred for 3 h, then diluted with in CH₂Cl₂and washed sequentially with water, saturated NaHCO₃ solution, andbrine. After drying over MgSO₄ and concentrating in vacuo, the mixturewas purified by chromatography on a silica gel column using 5% CH₃OH inCH₂Cl₂ as eluent to yield the title compound, a portion which waspurified by preparative reversed-phase C₁₈ HPLC using a linear gradientof 35% to 100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 36.0 mg99.0% pure compound. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC: Rt=16.45min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 188

Compound 188. A solution of 51 mg of the resulting compound of Example187A in 15 mL of methanol was hydrogenated under a slight positivepressure of hydrogen in the presence of 10 mg of 10% Pd(OH)₂ for 14 h.After filtering and concentrating in vacuo, the crude mixture was takenup into 10 mL CH₂Cl₂ and treated with 0.203 mL of DIEA and 19.0 mg of2-quinoxaloyl chloride. The mixture was stirred for 6 h, then dilutedwith CH₂Cl₂ and washed with water. After drying over MgSO₄ andconcentrating in vacuo, a portion of the mixture was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 2.1 mg of the titlecompound. TLC: Rf=0.25, 6% Ch₃CN/H₂O with 0.1% TFA for elution to obtain2.1 mg of the title compound. TLC: Rf=0.25, 6% CH₃OH in CH₂Cl₂. HPLC:Rt=16.21 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 189

-   A. Compound XXII (D′=isobutyl, A=H, E=4-acetamidophenyl,    trifluoroacetate salt). To a solution of 89.3 mg. (0.167 mmol) of    the resultant compound of Example 39B in CH₂Cl₂ (1 mL) at 0° to    50° C. was added trifluoromethanesulfonic acid (1 mL). After    stirring for 0.5 h the resultant mixture was concentrated in vacuo    and the resulting yellow gum used without subsequent purification.-   B. Compound 189. A solution of the resultant compound of Example    189A (0.167 mmol) in CH₂Cl₂ was treated sequentially, at ambient    temperature under an atmosphere of nitrogen, with 44.2 mg (0.217    mmol) of N-Boc-α-aminoisobutyric acid, 0.044 mL (0.251 mmol)    diisopropylethylamine, 27.1 mg (0.201 mmol) of    1-hydroxybenzotriazole hydrate, 38.5 mg (0.201 mmol)    1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The    mixture was stirred for 16 h and then concentrated in vacuo. The    residue was taken up in ethyl acetate and washed with water, 0.5 N    hydrochloric acid, washed with sodium bicarbonate, saturated brine,    dried over magnesium sulfate, filtered and concentrated in vacuo.    The residue was purified by low pressure silica gel column    chromatography using a 10% to 35% gradient of ethyl acetate/CH₂Cl₂    eluent to yield 69.3 mg of the title product as a white solid. TLC:    Rf=0.46, 60% ethyl acetate/CH₂Cl₂, HPLC: Rt=15.0 min; (¹H)-NMR    (CDCl₃) consistent with structure.

EXAMPLE 190

-   A. Compound XXXI (A=H, R³=methyl, R^(3′)=methyl, D′=isobutyl,    E=4-acetamidophenyl, hydrochloride salt). To a solution of 60.1 mg    of the resultant compound of Example 189B in CH₂Cl₂ (1 mL) at 0° to    5° C. was added trifluoromethanesulfonic acid (1 mL). After stirring    for 0.75 h, the resultant mixture was concentrated in vacuo and the    resulting white solid used directly for subsequent reaction.-   B. Compound 190. To a solution of 37 mg (0.059 mmol) of the    resultant compound of Example 190A in CH₂Cl₂ (3 mL) was added    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 15.4 mg (0.089 mmol) of 1-hydroxybenzotriazole    hydrate, and 17.8 mg (0.089 mmol) EDC. The mixture was stirred for    16 h and then concentrated in vacuo. The residue was taken up in    EtOAc and washed with saturated brine, dried over magnesium sulfate,    filtered and concentrated in vacuo. The residue was purified by thin    layer silica gel column chromatography using 50% of EtOAc in CH₂Cl₂    as eluent to yield 32.5 mg of the title product. TLC: Rf=0.35, 50%    EtOAc/CH₂Cl₂, HPLC: Rt=15.65 min; (¹H)-NMR (CDCl₃) consistent with    structure.

EXAMPLE 191

-   A. (2S, 3RS)-S-Amino-1-chloro-2-hydroxy-4-phenylbutane). A solution    of 2.24 g (6.71 mmol) of (1S,    2RS)-N-(1-benzyl-3-chloro-2-hydroxypropyl)-benzyloxycarbonylamine in    5 mL of methanol was added, at ambient temperature under a nitrogen    atmosphere, to a slurry of 0.22 g (10% by weight) of 10% palladium    on carbon in 60 mL methanol and hydrogenerated for 24 h, under a    slight positive pressure of hydrogen. The mixture was filtered and    concentrated in vacuo to yield 1.34 g of the mixed diastereomeric    products. TLC: Rf=0.33, 10% CH₃OH/CH₂Cl₂.-   B. (2S)-2-Benzyloxycarbonylamino-N¹-((1S,    2RS)-1-benzyl-3-chloro-2-hydroxypropyl)-N⁴-trityl succinamide. A    solution of 1.34 g (6.71 mmol) of the resultant compounds of Example    191A in 60 mL of dichloromethane was treated sequentially, at    ambient temperature under an atmosphere of nitrogen, with 3.58 g    (7.05 mmol) of Cbz-N^(δ)-trityl-asparagine, 0.95 g (7.05 mmol) of    1-hydroxybenzotriazole hydrate, 1.35 g (7.05 mmol) of EDC. The    mixture was stirred for 24 hours and then concentrated in vacuo. The    residue was taken up in ethyl acetate and washed with water,    saturated NaHCO₃, saturated NaCl; dried over MgSO₄; filtered and    concentrated in vacuo. The residue was purified by low pressure    silica gel column chromatography using 10% ethyl    acetate/dichloromethane as eluent to yield 3.08 g total of the mixed    diastereomeric products. TLC: Rf=0.75, 0.83, 40% EtOAc/CH₂Cl₂;    (¹H)-NMR (CDCl₃) consistent with structure.-   C. (2S)-2-Amino-N¹-((1S,    2RS)-1-benzyl-3-chloro-2-hydroxypropyl)-N⁴-trityl succinamide. A    solution of 2.80 g (4.06 mmol) of the resultant compounds of Example    191B in 5 mL of methanol was added, at ambient temperature under a    nitrogen atmosphere, to a slurry of 0.28 g (10% by weight) of 10%    palladium on carbon in 100 mL methanol and hydrogenated for 24 h    under a slight positive pressure of hydrogen. The mixture was    filtered and concentrated in vacuo to yield 2.26 g of the mixed    distereomeric products. TLC: Rf=0.42, 10% CH₃OH/CH₂Cl₂.-   D. (2S)-2-((1S,    2RS)-1-Benzyl-3-chloro-2-hydroxypropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-trityl    succinamide. A solution of 2.26 g (4.06 mmol) of the resultant    compounds of Example 191C in 60 mL of dichloromethane was treated    sequentially, at ambient temperature under an atmosphere of    nitrogen, with 0.74 g (4.27 mmol) of quinaldic acid, 0.58 g (4.27    mmol) of 1-hydroxybenzotriazole hydrate, and 0.82 g (4.27 mmol) of    EDC. After 24 hours, 30 mL of dichloromethane was added. The mixture    was washed with water, 5% NaHCO₃ solution, saturated NaCl, dried    over MgSO₄, filtered and concentrated in vacuo. The residue was    dissolved in 50% ethyl acetate/hexane and filtered through a plug of    silica gel. Removal of the solvents yielded 2.30 g of the mixed    diastereomeric products. TLC: Rf=0.53, 0.58, 40% EtOAc/CH₂Cl₂;    (¹H)-NMR (CDCl₃) consistent with structure.-   E.    (2S)-2-((1S,2RS)-1-Benzyl-2-hydroxy-3-iodopropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-trityl    succinamide. A solution of 1.05 g (1.48 mmol) of the resultant    compounds of Example 191D and 0.36 g (2.37 mmol) of sodium iodide in    15 mL of methyl ethyl ketone was heated to reflux for 24 hours. The    mixture was cooled to room temperature and then concentrated in    vacuo. The residue was taken up in dichloromethane and washed with    water, saturated NaCl, dried over MgSO₄, filtered and concentrated    in vacuo to yield 1.3 g of the mixed diastereomeric products. TLC:    Rf=0.58, 0.65, 40% EtOAc/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with    structure.-   F. (2S)-2-((1S, 2 syn,    anti)-3-(2-methylpropyl)amino-1-benzyl-2-hydroxypropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-trityl    succinamide. A solution of 207.6 mg (0.26 mmol) of the resultant    compounds of Example 191E and 0.5 mL (5.17 mmol) of isobutylamine in    9 mL of acetonitrile in a sealed tube was heated to reflux for 24    hours. After cooling to room temperature, the mixture was    concentrated in vacuo. The residue was taken up in dichloromethane    and washed with water, saturated NaCl, dried over MgSO₄, filtered    and concentrated in vacuo to yield 209.2 mg of the mixed    diastereomeric products. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂; (¹H)-NMR    (CDCl₃) consistent with structure.-   G. Compound XIV ((syn, anti)-OH, P=quinoline-2-carbonyl,    D′=isobutyl). A solution of 192.9 mg (0.26 mmol) of the resultant    compounds of Example 191F and 0.07 mL (0.388 mmol) of    diisopropylethylamine in 5 mL of dichloromethane was treated with    112.9 mg (0.517 mmol) of di-tert-butyldicarbonate. After 24 hours,    the mixture was diluted with dichloromethane. The mixture washed    with water, 5% NaHCO₃, 0.5 N HCl, saturated NaCl, dried over MgSO₄,    filtered and concentrated in vacuo. The residue was purified by low    pressure silica gel column chromatography using 40% ethyl    acetate/dichloromethane as eluent to yield 147.3 mg of the mixed    diastereomeric products. TLC: Rf=0.60, 0.67, 40% EtOAc/CH₂Cl₂;    (¹H)-NMR (CDCl₃) consistent with structure.-   H. Compounds 191. A solution of 147.3 mg (0.174 mmol) of the    resultant compounds of Example 191G in 2 mL of dichloromethane was    treated with 2 mL of trifluoroacetic acid. After 4 hours, the    mixture was concentrated in vacuo. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂.    To a solution of the resultant compound in 2 mL of dichloromethane    was sequentially added 0.5 mL of saturated NaHCO₃, small amount of    solid NaHCO₃ and 67 mg (0.226 mmol) of a mixture of    4-acetamido-3-fluorobenzenesulphonyl chloride and    3-acetamido-4-fluorobenzenesulphonyl chloride. After 3 hours, the    mixture was diluted with dichloromethane. The two layers were    separated and the aqueous layer was extracted once with    dichloromethane. The combined organic layer was washed with    saturated NaCl then dried over MgSO₄, filtered and concentrated in    vacuo. The residue was purified by low pressure silica gel column    chromatography using 2% methanol/dichloromethane was eluent to yield    64 mg of the mixed diastereomers and regioisomers which were further    purified with preparative HPLC to yield 18.9 mg of the mixed    regioisomers comprising compounds 191 as a white solid. TLC:    Rf=0.14, 5% CH₃OH/CH₂Cl₂; HPLC, Rt=13.36 min; (¹H)-NMR (CDCl₃)    consistent with structure.

EXAMPLE 193

Compound 193. A solution of 81.2 mg (0.096 mmol) of the resultant lowerRf diastereomer of Example 9/192A in 3 mL of dichloromethane was treatedwith 3 mL of trifluoroacetic acid. After 4 hours, the mixture wasconcentrated in vacuo. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solution of20.6 mg (0.0431 mmol) of the resultant residue in 1 mL ofdichloromethane was sequentially added 0.3 mL of saturated NaHCO₃, smallamount of solid NaHCO₃ and 12.4 mg (0.053 mmol) of4-acetamidobenzenesulphonyl chloride. After 3 hours, the mixture wasdiluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 8.3 mg of the title compound as a white solid; TLC: Rf=0.10, 5%CH₃OH/CH₂Cl₂; HPLC, Rt=12.7 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 194

Compound 194. To a solution of 13.0 mg (0.026 mmol) of thetrifluoroacetic acid deprotection product described in Example 193 in 1mL of dichloromethane was sequentially added 0.3 mL of saturated NaHCO₃,small amount of solid NaHCO₃ and 8.4 mg (0.033 mmol) of5-(isoxazol-3-yl)thiophene-2-sulphonyl chloride. After 3 hours, themixture was diluted with dichloromethane. The two layers were separatedand the aqueous layer was extracted once with dichloromethane. Thecombined organic layer was washed with brine then dried over MgSO₄,filtered and concentrated in vacuo. The residue was purified bypreparative HPLC to yield 5.1 mg of the title product as a white solid;TLC: Rf=0.27, 5% CH₃OH/CH₂Cl₂; HPLC, Rt=14.4 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 195

-   A. Compound XXII (A=(S)-3-tetrahydrofuryl, D′=cyclopentylmethyl,    A′=tert-butoxycarbonyl). To a solution of 264 mg of the resultant    compound of Example 140D in 10 mL of CH₂Cl₂ was added 0.14 mL of    disopropylethylamine and 175 mg of di-tert butylpyrocarbonate. After    stirring for 4 hours, the mixture was diluted with 50 mL of CH₂Cl₂,    washed with 0.5N of HCl and brine, dried over magnesium sulfate,    filtered and concentrated in vacuo to yield 364 mg of the title    compound as a white solid which was used without subsequent    purification. TLC: Rf=0.58, 40% EtOAc/CH₂Cl₂.-   B. A solution of 334 mg of the resultant compound of Example 195A in    5 mL of ethanol was hydrogenated under 30 psi of hydrogen in the    presence of 80 mg of platinum (IV) oxide for 24 hours. The mixture    was filtered and concentrated. The residue was purified by a low    pressure silica gel column chromatography using 20% EtOAc in CH₂Cl₂    eluent to yield 268 mg of the title compound. TLC: Rf=0.55, 40%    EtOAc/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.-   C. A solution of 268 mg of the resultant compound of Example 195B in    10 mL of EtOAc was treated with anhydrous HCl gas for 5 min. The    reaction mixture was sparged with nitrogen then concentrated in    vacuo and the resulting white solid used without subsequent    purification for subsequent reaction.-   D. Compound 195. To a solution of 233 mg of the crude resultant    compound of Example 195C in 10 mL of CH₂Cl₂ was added 2 mL of    saturated aqueous sodium bicarbonate and 149 mg of    4-methyloxybenzene sulfonyl chloride. After 3 hours, the resulting    mixture was diluted with CH₂Cl₂, washed with sodium bicarbonate,    brine, dried over magnesium sulfate, filtered and concentrated in    vacuo. The residue was purified by low pressure silica gel column    chromatography using 0% to 20% EtOAc/CH₂Cl₂ to yield 225 mg of the    title compound as a white solid. TLC: Rf=0.40, 20% EtOAc/CH₂Cl₂;    HPLC: Rt=15.65 min.: (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 196

-   A.    (1S,2S)-N-(1-Isobutyl-3-chloro-2-hydroxypropyl)benzyloxycarbonylamine.    To a solution of N-Cbz-leucine chloromethyl ketone (2.0 g) in 20 mL    of methanol was added, at 0° C., 1.0 g of sodium borohydride and the    mixture was stirred at ambient temperature for 24 h. The solution    was concentrated under reduced pressure and the residue partitioned    between 20 mL of saturated aqueous NH₄Cl and 500 ml of diethyl    ether. The organic fraction was separated, dried over MgSO₄ and    concentrated in vacuo and the residue purified by silica gel    chromatography to yield 1.8 g of white solid.-   B. (1S)-1-1(S)(Carbobenzyloxy)amino-2-isobutyl-oxirane. To a    solution of the resultant compound of Example 196A (300 mg) in    absolute ethanol was added 67 mg of powdered KOH. The mixture was    stirred for 3 h at ambient temperature, filtered through    diatomaceous earth, and concentrated in vacuo. The residue was    dissolved in diethyl ether, dried over MgSO₄, and concentrated to    yield 230 mg of colorless oil, which was used directly for    subsequent reaction.-   C.    (2R,3S)-N³-Carbobenzyloxy-N¹-isobutyl-1,3-diamino-2-hydroxy-5-methylhexane.    A 230 mg portion of the resultant compound of example 196B was    suspended in 5 mL of isobutylamine and the mixture stirred overnight    at ambient temperature. The mixture was concentrated in vacuo to    yield the title product as 179 mg of a white solid, which was used    directly for subsequent reaction.-   D. Compound I (A=tert-butoxycarbonyl, x=0, D=isobutyl,    E=4-methoxyphenyl, (s)-hydroxy). Following the procedure described    in Example 81, a solution of the resultant compound of example 196C    (170 mg) in CH₂Cl₂ was reacted with 4-methoxybenzenxulfoyl chloride    (150 mg) in the presence of aqueous NaHCO₃. Workup and silica gel    chromatography yielded 90 mg of product as a white solid.-   E. Compound I (A=H, x=0, D=isobutyl, E=4-methoxyphenyl,    (S)-hydroxy). A solution of the resultant compound of Example 196D    (90 mg) in ethanol was treated with 50 mg of 10% palladium on carbon    and the mixture stirred under an atmosphere of hydrogen. After    completion of reaction, the mixture was filtered and concentrated in    vacuo to yield 60 mg of the title compound which was used directly    for subsequent reaction.-   F. Compound 196. Reaction of the resultant compound of Example 196E    (60 mg) in CH₂Cl₂ was reacted with the resultant product of example    48A (150 mg) as described earlier yielded, following aqueous workup,    drying over MgSO₄, filtering, and concentration in vacuo, a residue    which was purified by silica gel chromatography using    methanol/CH₂CL₂ as eluant to yield the title product as 40 mg of    white solid. [¹h]-NMR(CDCl₃) consistent with structure.

EXAMPLE 197

We measured the inhibition constants of the compounds listed in TableVII against HIV-1 protease using the above-cited method of Pennington etal.

We also measured the anti-viral potency of the compounds in CCRM-CEMcells by the above-cited method of Meek et al. In the Tables below,K_(i) and IC₉₀ values are expressed in nM.

In Table VIII, the following classifications have been employed:

-   A: inhibits HIV replication at concentration of 100 nM or less.-   B: inhibits HIV replication at concentration of between 101 and    1,000 nM.-   C: inhibits HIV replication at a concentration of between 1,001 and    10,000 nM.-   D: inhibits HIV replication at a concentration of between 10,001 and    40,000 nM.-   ND: not tested.

TABLE VII Compound K_(i)value 1 4.0 2 2.0 3 32 4 19 5 2.0 6 3.0 7 8.0 8850 9 4.0 10 4.0 11 34 12 0.1 13 0.2 14 0.1 15 <0.1 16 <0.1 17 <0.1 18<0.1 19 <0.1 20 0.1 21 0.7 22 1.0 23 1.5 24 32,500 25 3,000 26 0.1 278.0 28 17 29 17 30 61 31 ND 32 2.5 33 80 34 17 35 4.0 36 19 37 0.1 381.5 39 17 40 1,100 41 220 42 46 43 4,200 44 5.0 45 6.0 46 154 47 4.0 481.4 49 9.0 50 11 51 ND 52 0.4 53 27 54 22 55 430 56 60 57 200 58 34 59206 60 4.0 61 4.0 62 72 63 7.0 64 3.0 65 0.7 66 0.4 67 7,400 68 120 6942 70 25 71 470 72 4000 73 140 74 11 75 290 76 ND 77 ND 78 ND 79 ND 80ND 81 2.3 82 1.5 83 ND 84 1.4 85 4.0 86 5.0 87 10 88 1.4 89 2.0 90 93 912.5 92 20 93 0.8 94 1.7 95 1.3 96 8.0 97 2.5 98 0.5 99 0.24 100 0.16 101250 102 33 103 4.5 104 5.5 105 7.5 106 1.4 107 1.4 108 2.0 109 6.0 11028 111 0.3 112 4.0 113 3.0 114 0.35 115 0.5 116 <0.1 117 0.26 118 <0.1119 1.8 120 11 121 2.0 122 1.2 123 10 124 1.1 125 0.3 126 310 127 650128 >5000 129 19 130 14 131 60 132 6.0 133 24 134 8.4 135 2.7 136 18 13726 138 1.4 139 1.2 140 <0.1 141 0.1 142 <0.1 143 <0.1 144 8.0 145 1.4146 2.0 147 1.6 148 0.2 149 1.7 150 6.0 151 0.8 152 2.5 153 0.2 154 0.5155 1.7 156 2.8 157 0.7 158 <0.1 159 0.2 160 1.0 161 20 162 0.5 163 0.5164 130 165 0.4 166 <0.1 167 0.45 168 0.6 169 <0.1 170 0.2 171 0.2 17221 173 0.6 174 10 175 0.1 176 <0.1 177 <0.1 178 0.1 179 0.4 180 <0.1 1810.3 182 0.2 183 0.1 184 5.0 185 3.5 186 140 187 0.3 188 11.5 189 5,500190 ND 191 33 192 67 193 400 194 350 195 0.2 196 ND 1001 220 1002 281003 8 1004 0.2 1005 250 1006 8 1007 42 1008 20 1009 20 1010 4 1011 15001012 11 1013 1500 1014 9300 1015 19

TABLE VIII Compound IC₉₀ Range 1 C 2 B 3 C 4 C 5 B 6 B 7 D 8 ND 9 B 10 B11 ND 12 A 13 A 14 A 15 A 16 B 17 B 18 B 19 B 20 A 21 A 22 B 23 B 24 ND25 ND 26 B 27 C 28 ND 29 C 30 ND 31 ND 32 C 33 ND 34 ND 35 B 36 ND 37 B38 C 39 C 40 ND 41 ND 42 ND 43 ND 44 B 45 C 46 ND 47 C 48 B 49 C 50 C 51C 52 B 53 ND 54 C 55 ND 56 ND 57 ND 58 ND 59 ND 60 C 61 C 62 ND 63 C 64C 65 C 66 B 67 ND 68 ND 69 ND 70 ND 71 ND 72 ND 73 ND 74 ND 75 ND 76 ND77 ND 78 ND 79 ND 80 ND 81 C 82 C 83 ND 84 C 85 C 86 B 87 C 88 B 89 C 90ND 91 B 92 ND 93 B 94 B 95 C 96 ND 97 B 98 B 99 B 100 A 101 ND 102 ND103 C 104 C 105 ND 106 C 107 C 108 C 109 B 110 ND 111 C 112 B 113 B 114B 115 B 116 A 117 C 118 B 119 C 120 ND 121 C 122 C 123 ND 124 D 125 B126 ND 127 ND 128 ND 129 ND 130 ND 131 ND 132 ND 133 ND 134 ND 135 C 136ND 137 ND 138 B 139 B 140 A 141 B 142 A 143 A 144 B 145 B 146 B 147 B148 A 149 B 150 B 151 C 152 ND 153 ND 154 ND 155 B 156 B 157 B 158 A 159B 160 A 161 ND 162 C 163 B 164 ND 165 B 166 A 167 B 168 A 169 A 170 B171 A 172 ND 173 A 174 ND 175 A 176 ND 177 ND 178 ND 179 ND 180 ND 181ND 182 B 183 B 184 ND 185 ND 186 ND 187 B 188 C 189 ND 190 ND 191 C 192C 193 ND 194 ND 195 A 196 ND 1001 ND 1002 ND 1003 B 1004 A 1005 ND 1006ND 1007 ND 1008 ND 1009 ND 1010 ND 1011 ND 1012 ND 1013 ND 1014 ND 1015A

As demonstrated in Tables VII and VIII, all of the compounds testeddisplayed inhibitory and anti-viral activity. Moreover, several of thesecompounds exhibited activity levels far greater than those of known HIVprotease inhibitors.

While we have described a number of embodiments of this invention, it isapparent that our basic constructions may be altered to provide otherembodiments which utilize the products and processes of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims, rather than by the specificembodiments which have been presented by way of example.

1. A compound of formula I:

wherein: A is selected from the group consisting of —R¹—C₁-C₆ alkyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, —NR²—CO—N(R²)(R²)and —CO—N(R²)(R²); each R¹ is independently selected from the groupconsisting of —C(O)—, —S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂,—NR²—S(O)₂—, —NR²—C(O)— and —NR²—C(O)—C(O)—; each R² is independentlyselected from the group consisting of H and C₁-C₃ alkyl optionallysubstituted with Ar; with the proviso that when R² is C₁-C₃ alkylsubstituted with Ar, said Ar may not be substituted with anAr-containing moiety; B, when present, is —N(R²)—C(R³)(R³)—C(O)—; x is 0or 1; each R³ is independently selected from the group consisting of H,Het, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆ cycloalkyl and C₅-C₆cycloalkenyl, wherein any member of said R³, except H, may be optionallysubstituted with one or more substituents selected from the groupconsisting of —OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Het, —CN, —SR²,—CO₂R², NR²—C(O)—R²; each n is independently 1 or 2; D and D′ areindependently selected from the group consisting of Ar; C₁-C₄ alkyl,which may be optionally substituted with one or more groups selectedfrom C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₂-C₄ alkenyl, which maybe optionally substituted with one or more groups selected from thegroup consisting of C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₃-C₆cycloalkyl, which may be optionally substituted with or fused with Ar;and C₅-C₆ cycloalkenyl, which may be optionally substituted with orfused with Ar; each Ar is independently selected from the groupconsisting of phenyl; 3-6 membered carbocyclic ring, wherein saidcarbocyclic ring may be saturated or unsaturated and optionallysubstituted with one or more groups selected from the group consistingof oxo, —OR², —R², —N(R²)(R²), —N(R²)—C(O)—R², C₁-C₃ alkyl substitutedwith —OH and optionally substituted with Ar, —CN, —CO₂R²,—C(O)—N(R²)(R²), halo and —CF₃; E is selected from the group consistingof Het; O-Het; Het-Het; —O—R³; —NR²R³; C₁-C₆ alkyl, which may beoptionally substituted with one or more groups selected from the groupconsisting of R⁴ and Het; C₂-C₆ alkenyl, which may be optionallysubstituted with one or more groups selected from the group consistingof R⁴ and Het; C₃-C₆ saturated carbocycle, which may optionally besubstituted with one or more groups selected from the group consistingof R⁴ and Het; and C₅-C₆ unsaturated carbocycle, which may optionally besubstituted with one or more groups selected from the group consistingof R⁴ and Het; each Het is independently selected from the groupconsisting of C₃-C₇ cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7membered saturated or unsaturated heterocycle, containing one heteroatomselected from N, N(R²), O, S and S(O)_(n), wherein said heterocycle mayoptionally be benzofused; and wherein any member of said Het may beoptionally substituted with one or more substituents selected from thegroup consisting of oxo, —OR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)(R²), —S(O)₂—N(R²)(R²), —N(R²)—C(O)—R₂, —C(O)—R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—Ar, methylenedioxy, —N(R²)—S(O)₂(R²),halo, —CF₃, —NO₂, Ar and —O—Ar; and each R⁴ is independently selectedfrom the group consisting of —OR², —C(O)—NHR², —S(O)₂—NHR², halo,—NR²—C(O)—R² and —CN.
 2. The compound according to claim 1,characterized in that said compound has the structure of formula XXII:

and A, D′ and E are defined as in claim
 1. 3. The compound according toclaim 1, characterized in that said compound has the structure offormula XXXI:

and A, R³, D′ and E are defined as in claim
 1. 4. A compound accordingto claim 1, wherein: A is selected from the group consisting of—R¹-C₁-C₆ alkyl, which may be optionally substituted with one or moregroups selected from the group consisting of hydroxy, C₁-C₄ alkoxy; eachR¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—CO—, —O—S(O)₂— and —NR²—S(O)₂—; each R² isindependently selected from the group consisting of H and C₁-C₃ alkyl;B, when present, is —NH—CH(R³)—C(O)—; x is 0 or 1; R³ is selected fromthe group consisting of Het, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆cycloalkyl and C₅-C₆ cycloalkenyl, wherein any member of said R³ may beoptionally substituted with one or more substituents selected from thegroup consisting of —OR², —C(O)—NH—R², —S(O)_(n)—N(R²)₂, Het and —CN; nis 1 or 2; D and D′ are independently selected from the group consistingof Ar; C₁-C₄ alkyl, which may be optionally substituted with C₃-C₆cycloalkyl or Ar; C₂-C₄ alkenyl, which may be optionally substitutedwith C₃-C₆ cycloalkyl or Ar; C₃-C₆ cycloalkyl, which may be optionallysubstituted or fused with Ar; and C₅-C₆ cycloalkenyl, which may beoptionally substituted or fused with Ar; Ar is selected from the groupconsisting of phenyl; 3-6 membered carbocyclic ring wherein saidcarbocyclic ring may be saturated or unsaturated and optionallysubstituted with one or more groups selected from the group consistingof oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)R², —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)₂, halo and —CF₃; E is selected from the group consisting ofHet; —O—R³; —NR²R⁵; C₁-C₆ alkyl, which may be optionally substitutedwith one or more R⁴ or Het; C₂-C₆ alkenyl, which may be optionallysubstituted with one or more R⁴ or Het; C₃-C₆ saturated carbocycle,which may optionally be substituted with one or more R⁴ or Het; andC₅-C₆ unsaturated carbocycle, which may optionally be substituted withone or more R⁴ or Het; each Het is independently selected from the groupconsisting of C₃-C₇ cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7membered saturated or unsaturated heterocycle, containing one heteroatomselected from N, O and S, which may optionally be benzofused; whereinany member of said Het may be optionally substituted with one or moresubstituents selected from the group consisting of oxo, —OR², —R²,—N(R²)₂, —R²—OH, —CN, —CO₂R², —C(O)—N(R²)₂ and —S(O)₂—N(R²)₂; each R⁴ isindependently selected from the group consisting of —OR², —C(O)—NHR²,—S(O)₂—NHR², halo and —CN; and each R⁵ is independently selected fromthe group consisting of H and R³.
 5. The compound according to claim 1,wherein: R³ is selected from the group consisting of C₁-C₆ alkyl, C₂-C₆alkenyl, C₅-C₆ cycloalkyl, C₅-C₆ cycloalkenyl and a 5-6 memberedsaturated or unsaturated heterocycle, wherein any member of said R³ mayoptionally be substituted with one or more substituents selected fromthe group consisting of —OR², —C(O)—NH—R², —S(O)_(n)N(R²)(R²), Het, —CN,—SR², —C(O)₂R², NR²—C(O)—R²; and D′ is selected from the groupconsisting of C₁-C₃ alkyl and C₃ alkenyl, wherein said alkyl or alkenylmay optionally be substituted with one or more groups selected from thegroup consisting of C₃-C₆ cycloalkyl, —OR², —O—Ar and Ar.
 6. Thecompound according to claim 1, wherein said compound has a molecularweight less than or equal to about 700 g/mol.
 7. A compound according toclaim 6, wherein said compound has a molecular weight less than or equalto about 600 g/mol.